Award details

Understanding the regulation of alkaloid biosynthesis in opium poppy and breeding new varieties

Reference	BB/K018809/1
Principal Investigator / Supervisor	Professor Ian Graham
Co-Investigators / Co-Supervisors	Dr Marcelo Kern, Dr Thilo Winzer
Institution	University of York
Department	Biology
Funding type	Research
Value (£)	1,194,061
Status	Completed
Type	Research Grant
Start date	01/10/2013
End date	30/06/2018
Duration	57 months

Abstract

Papaver somniferum (opium poppy) remains one of the richest sources of natural products for the pharmaceutical industry. We recently reported that opium poppy contains a cluster of ten genes encoding five different enzyme classes responsible for the biosynthesis of noscapine (Winzer et al, Science 2012, 336:1704-8), an antitumour alkaloid that is currently in clinical trials in the USA. Our discovery of the ten gene cluster in opium poppy allowed us to rapidly elucidate the main steps in the previously unknown noscapine biosynthetic pathway. In the current proposal we will use a combination of metabolite profiling and next generation sequencing to establish if related species that also produce noscapine have an equivalent gene cluster, allowing us to gain new understanding about the evolution of gene clusters in higher plants. We will also characterise those remaining steps in the biochemical pathway which are not fully understood using a combination of in-planta based Virus Induced Gene Silencing (VIGS) and in-vitro characterisation of enzymes obtained by heterologous expression. Major questions remain about the regulation of flux into the noscapine branch versus the morphinan branch of alkaloid biosynthesis and to address these we will use a combination of RNA sequencing, quantitative proteomics and isotope labelling followed by deep metabolite profiling to pinpoint the regulatory steps. In order to learn more about the control of flux and to develop new poppy varieties we will cross noscapine producing poppies into a number of mutant backgrounds that produce altered levels of morphinan alkaloids including morphine, codeine, thebaine and oripavine. This will result in new varieties producing novel combinations of opiate alkaloids that are required by the pharmaceutical industry. The research programme will therefore contribute both fundamental knowledge and improved sustainable feedstocks for the pharmaceutical industry.

Summary

Plants have evolved to produce a vast array of complex chemical structures to fight off attacks from herbivores and pathogens and to protect themselves from often hostile environments. These chemical structures also provide plants with medicinal properties that human civilisation has relied on for millennia. Opium poppy is one of the most important medicinal plants in the world and represents the largest single source of natural products used by the pharmaceutical industry. Opium poppies produce opiate alkaloids such as morphine and codeine, which are one of the main classes of painkiller drugs. The chemical structures of these opiate alkaloids are very complex and plants remain the best factory for their production. Opium poppy also produces hundreds of other alkaloids and these have been a source of several other drugs. Opium poppy is commercially grown using broad acre farming methods and the crop is harvested mechanically and shipped to factories for extraction of the high value chemicals. Pharmaceutical companies such as GlaxoSmithKline and Johnson & Johnson have their own production systems for production of opiate alkaloids from opium poppy. Our group in the Centre for Novel Agricultural Products at the University of York have been working with GlaxoSmithKline to develop new varieties of poppy that produce more of the morphinan alkaloids such as morphine and codeine. We have also been investigating how opium poppy produces noscapine, an anti-tumour alkaloid that stops human cells dividing. Noscapine has been used as a human cough suppressant for decades. Its effectiveness in tackling various forms of cancer has been demonstrated more recently, and early stage clinical trials are in progress in the USA. We particularly wanted to discover how noscapine is made in opium poppy as this would help us to breed new varieties that make more of it and also to gain insight into related molecules that may also have useful pharmaceutical activity. Our work led to a major breakthrough that was published recently in the leading journal Science (Winzer et al, Science 2012, 336:1704-8). By comparing opium poppy varieties which make noscapine with those that do not we discovered that the pathway for synthesis of noscapine is controlled by a complex cluster of ten genes encoding five different enzyme classes. This is the most complex gene cluster ever found in plants and provides invaluable insights into the process of gene duplication and re-organisation driving cluster evolution. This proposal builds on our exciting breakthrough and aims to establish if the gene cluster is also present in other related species that also make noscapine. This will provide new insight into the mechanisms and processes involved in gene cluster evolution. Elucidating details of the biochemical pathway we discovered for noscapine biosynthesis will make us better able to design strategies for improved production of noscapine and related molecules. Noscapine is produced by a separate branch of the alkaloid biosynthesis pathway to the one used to produce morphine and codeine. What regulates the flow of molecules into these different branches is not understood and we now have the tools and knowledge to address this important question for the first time. Answering these questions not only adds to our knowledge of the most important medicinal plant in terms of natural product feedstock for the pharmaceutical industry, it also provides us with the knowledge platform and tools to develop new varieties of opium poppy that are optimised for production of alkaloids such as noscapine, morphine and codeine. The final objective of this proposal is to use molecular breeding methods to develop new varieties of opium poppy with optimised levels of key opiate alkaloids for the benefit of the pharmaceutical industry and the UK industrial biotechnology sector.

Impact Summary

Who will benefit from this research? In addition to benefiting a broad group of academic researchers as detailed above, this research will also benefit a wider group including: 1. The pharmaceutical industry and in particular the industrial partner on this proposal, GlaxoSmithKline. 2. The Industrial Biotechnology sector in the UK - high value chemicals from plants are recognised as being an important component of this sector - and therefore the UK economy. 3. Policy makers who require examples of how the UK knowledge base can be used to attract new activities into the high value chemicals from plants /industrial biotechnology sector in the UK 4. The general public - the outputs of this work will lead to the sustainable production of feedstocks to support continued delivery of existing drugs and the production of new drugs that are currently under development. How will they benefit from this research? Global demand for the opiate alkaloid, noscapine, presently exceeds supply. Currently the global manufacture of noscapine is focused on its recovery as by-product from opium produced mainly in India. Opium supply has reduced over time due to inefficient production methods (lancing poppy capsules), diversion concerns and complex downstream processing requirements. The majority of major opiate alkaloids (morphine, codeine, thebaine and oripavine) are produced through broad acre farming of poppies followed by chemical processing occurring on the deseeded harvested dry poppy capsule (poppy straw). The production of a high yielding noscapine poppy that can be processed as poppy straw is the key to unlocking a new and more secure supply chain for noscapine. With noscapine and noscapine derivatives presently undergoing Phase 1 and Phase 2 clinical trials as a low toxicity anti-cancer agent there is a global need for the development of a new noscapine supply chain. In addition to a focus on noscapine, the GlaxoSmithKline (GSK) Opiates Business is presently pursuing a number of activities centred on a northern hemisphere base. Several of these projects have arisen from the strategic focus on new business opportunities for the Opiates Business. These are seen as priority business outcomes for GSK supported from the proposed R&D programme with the Centre for Novel Agricultural Products (CNAP) at the University of York: i) Development of a secure global poppy straw based supply chain for the economic production of the opiate alkaloid, noscapine ii) Northern hemisphere (UK & Europe) poppy crop trials and poppy crop sourcing options iii) Development of a supply chain for the production of the opiate alkaloid, noscapine iv) Chemical extraction of poppy actives and development of a European based opiate raw material supply v) Advancing the fundamental understanding of poppy plant biochemistry and plant breeding to increase commercial poppy productivity and identify new pharmaceutical product opportunities vi) Underpinning the GSK global opiate R&D footprint, providing indirect support for a planned technical transfer to the UK & Ireland of some opiate manufacture and the development of value added opiate products. The Department of Business Innovation and Skills has identified Industrial Biotechnology as having significant growth potential for the benefit of the UK economy over the next 15 years. Developing centres of excellence that facilitate specific aims such as high value chemicals from plant biomass for use in the pharmaceutical sector has been identified in the roadmap as necessary to fulfil the potential of Industrial Biotechnology in the UK. The outputs of this research programme will contribute significantly to such aims by facilitating the development of industrial biotechnology to the benefit of the UK economy and the economic competitiveness of the UK in this important growth area.

Committee	Research Committee B (Plants, microbes, food & sustainability)
Research Topics	Crop Science, Pharmaceuticals, Plant Science
Research Priority	X – Research Priority information not available
Research Initiative	X - not in an Initiative
Funding Scheme	Industrial Partnership Award (IPA)

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