Cargando…
Enhancement of Inhibitory Activity by Combining Allosteric Inhibitors Putatively Binding to Different Allosteric Sites on Cathepsin K
Background: Cathepsin K, which is involved in bone resorption, is a good target for treating osteoporosis, but no clinically approved medicine has been developed. Recently, allosteric inhibitors with high specificity and few side effects have been attracting attention for use in new medicines. Metho...
Autores principales: | , , , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2023
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10221639/ https://www.ncbi.nlm.nih.gov/pubmed/37241936 http://dx.doi.org/10.3390/molecules28104197 |
_version_ | 1785049504417841152 |
---|---|
author | Sato, Shun Yamamoto, Kana Ito, Moeno Nishino, Katsutoshi Otsuka, Takanao Irie, Kazuhiro Nagao, Masaya |
author_facet | Sato, Shun Yamamoto, Kana Ito, Moeno Nishino, Katsutoshi Otsuka, Takanao Irie, Kazuhiro Nagao, Masaya |
author_sort | Sato, Shun |
collection | PubMed |
description | Background: Cathepsin K, which is involved in bone resorption, is a good target for treating osteoporosis, but no clinically approved medicine has been developed. Recently, allosteric inhibitors with high specificity and few side effects have been attracting attention for use in new medicines. Methods: Cathepsin K inhibitors were isolated from the methanol extract of Chamaecrista nomame (Leguminosae) using cathepsin K inhibition activity-assisted multi-step chromatography. Standard kinetic analysis was employed to examine the mechanism of cathepsin K inhibition when an isolated inhibitor and its derivative were used. The allosteric binding of these cathepsin K inhibitors was supported by a docking study using AutoDock vina. Combinations of allosteric cathepsin K inhibitors expected to bind to different allosteric sites were examined by means of cathepsin K inhibition assay. Results: Two types of cathepsin K inhibitors were identified in the methanol extract of Chamaecrista nomame. One type consisted of cassiaoccidentalin B and torachrysone 8-β-gentiobioside, and inhibited both cathepsin K and B with similar inhibitory potential, while the other type of inhibitor consisted of pheophytin a, and inhibited cathepsin K but not cathepsin B, suggesting that pheophytin a binds to an allosteric site of cathepsin K. Kinetic analysis of inhibitory activity suggested that pheophytin a and its derivative, pheophorbide b, bind allosterically to cathepsin K. This possibility was supported by a docking study on cathepsin K. The cathepsin K inhibitory activity of pheophytin a and pheophorbide b was enhanced by combining them with the allosteric inhibitors NSC 13345 and NSC94914, which bind to other allosteric sites on cathepsin K. Conclusions: Different allosteric inhibitors that bind to different sites in combination, as shown in this study, may be useful for designing new allosteric inhibitory drugs with high specificity and few side effects. |
format | Online Article Text |
id | pubmed-10221639 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2023 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
spelling | pubmed-102216392023-05-28 Enhancement of Inhibitory Activity by Combining Allosteric Inhibitors Putatively Binding to Different Allosteric Sites on Cathepsin K Sato, Shun Yamamoto, Kana Ito, Moeno Nishino, Katsutoshi Otsuka, Takanao Irie, Kazuhiro Nagao, Masaya Molecules Article Background: Cathepsin K, which is involved in bone resorption, is a good target for treating osteoporosis, but no clinically approved medicine has been developed. Recently, allosteric inhibitors with high specificity and few side effects have been attracting attention for use in new medicines. Methods: Cathepsin K inhibitors were isolated from the methanol extract of Chamaecrista nomame (Leguminosae) using cathepsin K inhibition activity-assisted multi-step chromatography. Standard kinetic analysis was employed to examine the mechanism of cathepsin K inhibition when an isolated inhibitor and its derivative were used. The allosteric binding of these cathepsin K inhibitors was supported by a docking study using AutoDock vina. Combinations of allosteric cathepsin K inhibitors expected to bind to different allosteric sites were examined by means of cathepsin K inhibition assay. Results: Two types of cathepsin K inhibitors were identified in the methanol extract of Chamaecrista nomame. One type consisted of cassiaoccidentalin B and torachrysone 8-β-gentiobioside, and inhibited both cathepsin K and B with similar inhibitory potential, while the other type of inhibitor consisted of pheophytin a, and inhibited cathepsin K but not cathepsin B, suggesting that pheophytin a binds to an allosteric site of cathepsin K. Kinetic analysis of inhibitory activity suggested that pheophytin a and its derivative, pheophorbide b, bind allosterically to cathepsin K. This possibility was supported by a docking study on cathepsin K. The cathepsin K inhibitory activity of pheophytin a and pheophorbide b was enhanced by combining them with the allosteric inhibitors NSC 13345 and NSC94914, which bind to other allosteric sites on cathepsin K. Conclusions: Different allosteric inhibitors that bind to different sites in combination, as shown in this study, may be useful for designing new allosteric inhibitory drugs with high specificity and few side effects. MDPI 2023-05-19 /pmc/articles/PMC10221639/ /pubmed/37241936 http://dx.doi.org/10.3390/molecules28104197 Text en © 2023 by the authors. https://creativecommons.org/licenses/by/4.0/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 (https://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Article Sato, Shun Yamamoto, Kana Ito, Moeno Nishino, Katsutoshi Otsuka, Takanao Irie, Kazuhiro Nagao, Masaya Enhancement of Inhibitory Activity by Combining Allosteric Inhibitors Putatively Binding to Different Allosteric Sites on Cathepsin K |
title | Enhancement of Inhibitory Activity by Combining Allosteric Inhibitors Putatively Binding to Different Allosteric Sites on Cathepsin K |
title_full | Enhancement of Inhibitory Activity by Combining Allosteric Inhibitors Putatively Binding to Different Allosteric Sites on Cathepsin K |
title_fullStr | Enhancement of Inhibitory Activity by Combining Allosteric Inhibitors Putatively Binding to Different Allosteric Sites on Cathepsin K |
title_full_unstemmed | Enhancement of Inhibitory Activity by Combining Allosteric Inhibitors Putatively Binding to Different Allosteric Sites on Cathepsin K |
title_short | Enhancement of Inhibitory Activity by Combining Allosteric Inhibitors Putatively Binding to Different Allosteric Sites on Cathepsin K |
title_sort | enhancement of inhibitory activity by combining allosteric inhibitors putatively binding to different allosteric sites on cathepsin k |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10221639/ https://www.ncbi.nlm.nih.gov/pubmed/37241936 http://dx.doi.org/10.3390/molecules28104197 |
work_keys_str_mv | AT satoshun enhancementofinhibitoryactivitybycombiningallostericinhibitorsputativelybindingtodifferentallostericsitesoncathepsink AT yamamotokana enhancementofinhibitoryactivitybycombiningallostericinhibitorsputativelybindingtodifferentallostericsitesoncathepsink AT itomoeno enhancementofinhibitoryactivitybycombiningallostericinhibitorsputativelybindingtodifferentallostericsitesoncathepsink AT nishinokatsutoshi enhancementofinhibitoryactivitybycombiningallostericinhibitorsputativelybindingtodifferentallostericsitesoncathepsink AT otsukatakanao enhancementofinhibitoryactivitybycombiningallostericinhibitorsputativelybindingtodifferentallostericsitesoncathepsink AT iriekazuhiro enhancementofinhibitoryactivitybycombiningallostericinhibitorsputativelybindingtodifferentallostericsitesoncathepsink AT nagaomasaya enhancementofinhibitoryactivitybycombiningallostericinhibitorsputativelybindingtodifferentallostericsitesoncathepsink |