Cargando…

Optimization of the production process for the anticancer lead compound illudin M: downstream processing

BACKGROUND: Secondary metabolites have played a key role as starting points for drug development programs due to their often unique features compared with synthetically derived molecules. However, limitations related to the discovery and supply of these molecules by biotechnological means led to the...

Descripción completa

Detalles Bibliográficos
Autores principales: Chaverra-Muñoz, Lillibeth, Briem, Theresa, Hüttel, Stephan
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9382783/
https://www.ncbi.nlm.nih.gov/pubmed/35978411
http://dx.doi.org/10.1186/s12934-022-01886-2
_version_ 1784769356188614656
author Chaverra-Muñoz, Lillibeth
Briem, Theresa
Hüttel, Stephan
author_facet Chaverra-Muñoz, Lillibeth
Briem, Theresa
Hüttel, Stephan
author_sort Chaverra-Muñoz, Lillibeth
collection PubMed
description BACKGROUND: Secondary metabolites have played a key role as starting points for drug development programs due to their often unique features compared with synthetically derived molecules. However, limitations related to the discovery and supply of these molecules by biotechnological means led to the retraction of big pharmaceutical companies from this field. The reasons included problems associated with strain culturing, screening, re-discovery, purification and characterization of novel molecules from natural sources. Nevertheless, recent reports have described technical developments that tackle such issues. While many of these reports focus on the identification and characterization of such molecules to enable subsequent chemical synthesis, a biotechnological supply strategy is rarely reported. This may be because production processes usually fall under proprietary research and/or few processes may meet the requirements of a pharmaceutical development campaign. We aimed to bridge this gap for illudin M—a fungal sesquiterpene used for the development of anticancer agents—with the intention to show that biotechnology can be a vital alternative to synthetic processes dealing with small molecules. RESULTS: We used µL-scale models to develop an adsorption and extraction strategy for illudin M recovery from culture supernatant of Omphalotus nidiformis and these findings were successfully transferred into lab-scale. By adsorbing and eluting the product using a fixed resin-bed we reduced the working volume by ~ 90% and removed the aqueous phase from the process. After a washing step, a highly concentrated illudin M fraction was obtained by isocratic elution with 80% methanol. The fraction was dried and extracted using a water/heptane mixture, enriching illudin M in the heptane phase. From heptane illudin M could be instantly crystalized by concentrating the solution, achieving a final purity > 95%. CONCLUSION: We have developed a robust, scalable and low-cost downstream process to obtain highly pure illudin M. By using solid phase extraction we reduced the production of solvent waste. Heptane from the final purification step could be recycled. The reduced amounts of solvents required, and the short purification time render this method a very economic and ecologic alternative to published processes. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s12934-022-01886-2.
format Online
Article
Text
id pubmed-9382783
institution National Center for Biotechnology Information
language English
publishDate 2022
publisher BioMed Central
record_format MEDLINE/PubMed
spelling pubmed-93827832022-08-18 Optimization of the production process for the anticancer lead compound illudin M: downstream processing Chaverra-Muñoz, Lillibeth Briem, Theresa Hüttel, Stephan Microb Cell Fact Methodology BACKGROUND: Secondary metabolites have played a key role as starting points for drug development programs due to their often unique features compared with synthetically derived molecules. However, limitations related to the discovery and supply of these molecules by biotechnological means led to the retraction of big pharmaceutical companies from this field. The reasons included problems associated with strain culturing, screening, re-discovery, purification and characterization of novel molecules from natural sources. Nevertheless, recent reports have described technical developments that tackle such issues. While many of these reports focus on the identification and characterization of such molecules to enable subsequent chemical synthesis, a biotechnological supply strategy is rarely reported. This may be because production processes usually fall under proprietary research and/or few processes may meet the requirements of a pharmaceutical development campaign. We aimed to bridge this gap for illudin M—a fungal sesquiterpene used for the development of anticancer agents—with the intention to show that biotechnology can be a vital alternative to synthetic processes dealing with small molecules. RESULTS: We used µL-scale models to develop an adsorption and extraction strategy for illudin M recovery from culture supernatant of Omphalotus nidiformis and these findings were successfully transferred into lab-scale. By adsorbing and eluting the product using a fixed resin-bed we reduced the working volume by ~ 90% and removed the aqueous phase from the process. After a washing step, a highly concentrated illudin M fraction was obtained by isocratic elution with 80% methanol. The fraction was dried and extracted using a water/heptane mixture, enriching illudin M in the heptane phase. From heptane illudin M could be instantly crystalized by concentrating the solution, achieving a final purity > 95%. CONCLUSION: We have developed a robust, scalable and low-cost downstream process to obtain highly pure illudin M. By using solid phase extraction we reduced the production of solvent waste. Heptane from the final purification step could be recycled. The reduced amounts of solvents required, and the short purification time render this method a very economic and ecologic alternative to published processes. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s12934-022-01886-2. BioMed Central 2022-08-17 /pmc/articles/PMC9382783/ /pubmed/35978411 http://dx.doi.org/10.1186/s12934-022-01886-2 Text en © The Author(s) 2022 https://creativecommons.org/licenses/by/4.0/Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) . The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/ (https://creativecommons.org/publicdomain/zero/1.0/) ) applies to the data made available in this article, unless otherwise stated in a credit line to the data.
spellingShingle Methodology
Chaverra-Muñoz, Lillibeth
Briem, Theresa
Hüttel, Stephan
Optimization of the production process for the anticancer lead compound illudin M: downstream processing
title Optimization of the production process for the anticancer lead compound illudin M: downstream processing
title_full Optimization of the production process for the anticancer lead compound illudin M: downstream processing
title_fullStr Optimization of the production process for the anticancer lead compound illudin M: downstream processing
title_full_unstemmed Optimization of the production process for the anticancer lead compound illudin M: downstream processing
title_short Optimization of the production process for the anticancer lead compound illudin M: downstream processing
title_sort optimization of the production process for the anticancer lead compound illudin m: downstream processing
topic Methodology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9382783/
https://www.ncbi.nlm.nih.gov/pubmed/35978411
http://dx.doi.org/10.1186/s12934-022-01886-2
work_keys_str_mv AT chaverramunozlillibeth optimizationoftheproductionprocessfortheanticancerleadcompoundilludinmdownstreamprocessing
AT briemtheresa optimizationoftheproductionprocessfortheanticancerleadcompoundilludinmdownstreamprocessing
AT huttelstephan optimizationoftheproductionprocessfortheanticancerleadcompoundilludinmdownstreamprocessing