A molecular dissection of the interplay between diabetes and cancer: an integrated, multidisciplinary approach

Lead Research Organisation: University of York
Department Name: Chemistry

Abstract

This research combines molecular science, cell biology and disease processes - it is in molecular medicine. The research programme focuses on human insulin and Insulin-like Growth Factors I and II (IGF-I/II); these are closely related protein hormones. Their separate evolution has resulted in their acquiring separate biological functions, with insulin becoming a key regulator of metabolism, while IGF-I/II are major growth factors. When released into the blood the hormones bind, tightly and specifically, to their receptors, (Insulin Receptor (IR) and IGF-1R respectively); these are large complex protein molecules on the cell surface. Receptor binding, through which the hormone activity is expressed, involves structural changes in both the hormone and the receptors.

Despite their fundamental medical importance (insulin: in diabetes, IGFs: in development, cancer, aging) the active conformations of the hormones in their hormone:receptor complexes are not known. Thus, more than 40 years after the first insulin crystal structure was determined, all the analogues used in treatment of diabetes are still based on the inactive/storage forms of this hormone.

Insulin cross-reation with IGF-1R and its accumulation in Type 2 Diabetes entails this hormone in cell proliferation and growth. As IGF-I/II are also cancer-specific growth factors the identification of pure 'metabolic' and 'cell growth' structural signatures of insulin and IGF-I/II is of fundamental importance not only for the understanding of the biology of these hormones, but also for new, effective treatments of diabetes and cancer. Therefore this Programme is a response to an urgent need for a unique, consolidated, and multidisciplinary attack on these critical problems of insulin and IGF-I/II structure and biology.

This programme combines fundamental (structural biology, cell signaling, organic and protein chemistry) and applied research. The fundamental research provides the foundations of the programme. However, advanced expertise of this group in the applied biomedical sciences will enable parallel pursuit of applied aims as well.
This programme will deliver: (i) 3-D structural description of insulin:IR complexes; elucidation of the active form of human insulin, (ii) identification of active surfaces in insulin's responsible for metabolic effects, (iii) development of novel, specific metabolic insulin analogues, (iv) identification of the mitogenic structural signatures of the Insulin-like Growth Factors-I/II (IGF-I/II) expressed through IR, (vi) initiation of developing organo/peptido-mimetics of these hormones.

We believe that the potential impacts on molecular cell biology and on medical science are immense as it addresses directly two major health problems, diabetes and cancer. Both have enormous social and economic consequences. (i) Firstly, solution of the central problem of diabetes: insulin-Insulin Receptor (IR) interaction, i.e. activation of insulin from its storage form to its IR stimulating conformation, will provide a long-awaited breakthrough in understanding of insulin structure-function relationships and open new possibilities for rational design of novel insulins. (ii) Subsequently, identification of structural determinants of insulin and IGF-I/II that govern their specific signaling pathways and which are responsible for metabolic and growth specificity of these hormones should enhance the prospects for rational design and production of pure and safe insulins without their undesired mitogenic properties. This is a problem indicated by some currently used clinical analogues. (iii) The studies should allow initiation of rational design and synthesis of IGF-I/II-specific antagonists for novel anti-cancer therapies. (iv) The firm control on hormone chemistry within this programme will also allow initiation of rational design and synthesis of organo-mimics of insulin, which may lead to insulin-like drugs suitable for oral delivery.

Technical Summary

The major experimental goals are the production of the IR construct (in a large-scale mammalian cell expression facility) and its crystallization with selected high affinity analogues. This will be followed by their X-ray crystallographic analysis. This research in turn, rests on chemical semi- and full-synthesis of insulin and IGF analogues and their structural and biological characterization, especially their binding affinity.

Crystals of IR constructs, one complexed with insulin and another with a high affinity analogue, have already been prepared. The immediate goal is now to improve the diffracting power of these crystals (currently at ~ 4.5 - 3.9 Å). The range of insulin analogues will be exploited and alternative receptors investigated if necessary. X-ray data collection will be undertaken at synchrotrons.
The structural changes in insulin and the receptor associated with complex formation will be analysed and compared to the native hormone's solution structure. With this information another cycle of insulin analogue synthesis and analysis will be undertaken. Experiments will be carried out to explore the functional roles of residues on insulin's receptor-binding surface. This work will be paralleled by synthesis of new insulins with novel chemistry. Insulin and IGF-I/II hybrids will also be synthetised and their structures (alone and in complex with IR) determined to give insight into the structural basis of the different specificity of these homones. Cell signalling profiles of all analogues will be monitored.

The programme involves four centres: University of York (YSBL) - structural biology, Institute of Organic Chemistry and Biochemistry (IOCB, Prague) - chemical biology, organic and protein chemistry, insulin affinity/potency essays, drug design/synthesis, University College Cork (UCC) - cell biology/signalling, Walter and Eliza Hall Institute of Medical Research, Melbourne (WEHI) - molecular and structural biology of IR.

Planned Impact

The research on insulin and its receptor proposed in this Programme will have a fundamental impact on understanding insulin's mechanism of action - and that of other related hormones/factors. Despite 40 years of intensive structural studies we still do not know the structure of the active form of insulin although we know that it must be different from its well-characterized storage and solution forms. Nor is an accurate structure of the insulin:Insulin Receptor complex known. With insight into insulin's structure and its contacts in its receptor- bound/active conformation, the nature of the hormone fold, the invariance of its amino acids distribution will be understood properly - at last. Most importantly, the design of new insulin analogues with new structures and properties will be possible.

Insulin and the closely related hormones IGF-I and IGF-II interact with each others receptors, this implies that insulin is not only involved in metabolic regulation but also in cell proliferation and growth. The active forms of IGF-I and II are also unknown. Progress in novel clinical approaches to diabetes and cancer is hindered by ignorance of the specific structural signatures in insulin and IGF-I/II responsible for their different metabolic and growth signaling properties. The proposed research programme is directly relevant to two major human diseases: diabetes and cancer, both of which present huge social and economic burdens. The "epidemic"-like scale of diabetes in particular presents an urgent and a long-term problem.
The importance of this research, and its urgency, is highlighted by the increase in certain forms of cancer (e.g. colorectal, breast) in Type 2 Diabetes, and a >50% increase in cardiovascular disease, stroke and kidney disease, and long-term complications directly linked to diabetes.

The main Programme's impact will come from:

(i) Understanding the structures involved in insulin-Insulin Receptor (IR) interaction, i.e. the activation of insulin from its storage/solution form to its IR stimulating conformation, will be described for the first time in 3-D detail. This will provide a breakthrough in understanding insulin structure-function relationships and open new possibilities for rational design of novel insulins by non-commercial and Pharmaceutical laboratories.

(ii) Subsequently, structures of more novel insulin analogues and their hybrids with IGF-I/II (both free and in complexes with IR) will be determined and their functions/functional effects will be characterized. This molecular approach should enable identification of structural determinants of insulin and IGF-I/II that govern their specific signaling pathways and which are responsible for the metabolic and growth specificity exhibited by these modified hormones. The results will extend the models available for more rational design and for production of safe insulins with only specific metabolic functions; we expect strong interest from the pharmaceutical industry in exploiting these discoveries.

(iii) The new structural information on the 'active' structures will also guide rational design and synthesis of insulin organo-mimics in this programme that may lead to new approaches in development of insulin-like drugs suitable for oral delivery. We anticipate that the new basis for rational drug design provided by the structures of the hormones/analogues in their receptor complexes will stimulate pharmaceutical research and development of oral treatments for diabetes and cancer.

Publications

10 25 50

 
Description Grant Reviewes for Diabetes UK
Geographic Reach National 
Policy Influence Type Participation in a advisory committee
 
Description BioProNET
Amount £99,089 (GBP)
Funding ID PoC Nov15 Brzozowski 
Organisation Biotechnology and Biological Sciences Research Council (BBSRC) 
Sector Public
Country United Kingdom
Start 04/2016 
End 03/2017
 
Description Diabetes UK Equipment Grant
Amount £39,034 (GBP)
Funding ID 14/0004972 
Organisation Abbott 
Department Abbott UK
Sector Private
Country United Kingdom
Start 11/2014 
 
Description Guy Dodson Fund
Amount £150,000 (GBP)
Organisation Novo Nordisk 
Sector Public
Country Denmark
Start 10/2014 
End 09/2018
 
Description PhD Studentship
Amount £69,400 (GBP)
Organisation Novo Nordisk 
Sector Public
Country Denmark
Start 10/2014 
End 09/2018
 
Description Research Priming Fund
Amount £17,000 (GBP)
Organisation University of York 
Sector Academic/University
Country United Kingdom
Start 08/2013 
End 07/2014
 
Title Development of novel mammalian cell-based expression system for hard-to-produce proteins 
Description This development aims to address some of the current challenges faced in the large-scale production of proteins, in particular (a) making the production of difficult to express proteins more cost and time-efficient, (b) providing an alternative to the existing eukaryotic expression systems when unsuccessful. Along with applying fundamental academic research to address industry challenges this application is aimed at providing a base upon which further, and larger, collaborative projects can be applied for. It bridges novel higher-risk fundamental research, difficult in an industrial/pharma environment, with multi-target bio-medical industrial applicability. The key to this approach is to use an existing system of intracellular storage bodies, known as Weibel-Palade bodies that have a crucial role in the storage and controlled secretion of haemostatic proteins. Our intention is to show that we can hijack this already nature-tested packaging system and use it to form part of a novel protein expression system. The main applied aim of this development is to plug in it into our current MRC Insulin Receptor-related project. 
Type Of Material Model of mechanisms or symptoms - in vitro 
Provided To Others? No  
Impact This project just has been awarded by BBSRC BioProNET network for one year (£99K). It will start 1 April 2016. The post-doc has already been recruited. 
URL http://biopronetuk.org/category/success-stories/
 
Title Translational tools to aid diagnosis and treatment of Type-2 Diabetes: Development of cell-based platforms to dissect insulin-signaling pathways. 
Description No human cell lines are currently available to assay specific, IR/IGF-1R insulin dependent insulin-stimulated GLUT4 trafficking. These are essential for to increase the translational capacity of this field. The cell lines generated here will therefore be of great and fundamental benefit to the Diabetes research community by: (i) provision of molecular detail about the build-up of insulin resistance in diabetes, (ii) translation of these findings into clinical applications by screening library of insulin analogues, (iii) opening a great collaboration potential between the York and others with interests in the field. 
Type Of Material Cell line 
Provided To Others? No  
Impact This is work in progress; will also be a part of the MRC Programme Grant renewal application. 
 
Title Insulin structure-function database 
Description A preliminary beta version of Insulin Database (ID) has been developed. This database integrates all known insulin's structures, its mutations, and functionality. It is in a working but preliminary form. Funds/grants will be considered for further development and maintenance of this database. 
Type Of Material Database/Collection of data 
Provided To Others? No  
Impact Progress in insulin/diabetes field is tampered by lack of consolidated, unified and user-friendly insulin database that could be used by a wide audience: researchers, clinicians, students, college students and non-professional public. This database should make all insulin analogues and mutants 'easy' to understand and compare even by a lay person. Beside its 'popular' and educational characters it is, first of all, a professional tool in studying and working with insulin on clinical and very advanced research level. 
 
Description Collaboration with Prof. Nia Bryant - Dept. of Biology, University of York - Translational tools to aid diagnosis and treatment of Type-2 Diabetes. 
Organisation University of York
Department Department of Computer Science
Country United Kingdom 
Sector Academic/University 
PI Contribution There is currently a major knowledge gap in the understanding of the downstream effects of activation of different insulin receptors. This gap is evident in the studies of insulin-induced GLUT4 trafficking, which typically use different cell types without determining which IR isoforms are present. By combining the strong track records in GLUT4 (N Bryant) and IR/insulin/IGF research (AM Brzozowski), this project aims to generate tools to remedy this and to pave the way for careful, analytical studies of molecular aspects of insulin action, bringing to diabetes-related research a new level of molecular sophistication and control.
Collaborator Contribution Generation of human cells with controlled expression of glucose transporter and insulin receptor.
Impact Research output (still confidential) that will be an integral part of MRC Programme Grant renewal application.
Start Year 2016
 
Description Main Collaborations within the MRC Programme grant 
Organisation Novo Nordisk
Country Denmark 
Sector Public 
PI Contribution The first year of this Programme involved collaborations between molecular/structural biology laboratory (York Structural Biology Laboratory (YSBL)- University of York), organic/protein chemistry (Institute of Organic Chemistry and Biochemistry Czech Academy of Sciences (IOCB) - Prague), cell biology (University College Cork (UCC)). The Walter and Eliza Hall Institute of Medical Research ((WEHI), Melbourne) deliver the know-how in large-scale of production of human insulin receptor (IR). All groups worked within the scope of the planned programme. YSBL work underpinned all other activities, focusing on development of sustainable and novel production of IR that is one of the major bottlenecks of insulin-related research. The Ysbl Eukarytioc Expression Facility (YEF) has been established in this process. It is self-sufficient for baculovirus and mammalian cloning and expression work for this project. Over 50 IR constructs have been generated, and expression of some is optimized. IOCB developed novel synthetic routes for generation of new insulin and IGF analogues. They also established binding and signaling essays for all IR isoforms, which serve the characterization and preliminary evaluation of the most promising analogues from steadily growing (>100) library of semi/fully-synthetic insulins generated in this project. More complex signaling networks have been studied further in the UCC. The first, large batch of IR for structural studies has also been provided by WEHI. Therefore the first year of this programme cemented its foundations and positively validated its organizational and scientific principles. The multifaceted character of this programme grant has already generated growing network of new collaborations. Notably, an agreement between the YSBL and Novo Nordisk A/S (Copenhagen) has been signed upon which a fund in memory of late Prof Guy Dodson has been established. £150K will be provided here by Novo-Nordisk towards the PhD studentships devoted to insulin-related research. The first PhD programme - closely linked with this MRC grant - will start in October 2014. This YSBL-Novo Nordisk PhD-based initiative generates an excellent and exciting opportunity to elevate the collaboration between these institutions into the field of diabetes research that will transgress existing limiting boundaries. Collaboration between Prof AM Brzozowski (YSBL) and Prof Pierre de Meyts (Professor Emeritus in the Dept. of Chemistry of Catholic University of Louvain (Belgium), ex-Director of Research of the Hagedorn Research Institute, Gentofte, DK)) has also been established in the field of insulin-related structural and cellular biology in insects. This aims to develop a new programme - tangential and complementary to this MRC award - that will be focused on insects' insulins (DLPs), IR and DLPs-binding proteins. This research - due to a remarkable similarity of human and insects' insulin-related biology - will provide an additional insight into specificity and structure-function relationships of insulins and IRs. It should also shed light on age-regulatory role of insulin and IGFs in humans. Several proteins have already been produced here and their complexes characterized by X-ray crystallography. Branching of this research into insects-based model of insulin function resulted also in a startup of collaboration between Prof AM Brzozowski (YSBL) and Dr Sean T Sweeney (Dept. of Biology, University of York). This should expand the studies on biology of insects' insulins/IR systems (initiated with Prof P de Meyts) into probing of the phenotype modulation by insulin-signaling related genes knock-outs/rescue-based studies. They may lead to insect-based models for diabetes and life-span regulation in humans. Dr Sweeney has already developed key tools for studies of synaptic transmission and growth in Drosophila that delivered important insight into some forms of human neurodegenerative diseases, and molecular bases of aging and pain. These molecular tools will be applied for insulin-related signaling and regulatory systems. The second year of this Programme Grant resulted in cementing of York collaboration with Novo Nordisk (Copenhagen)(NN). Series of meetings in York and Copenhagen yielded detail definition of the expansion of Insulin Receptor (IR) research programme that is the core of this award. The agreement about collaboration, including details of the intellectual properties, publication rights, patenting etc. has been signed by both sides. Most advanced methodology in IR characterisation (developed in Copenhagen) has been made available to York group. Members of the YSBL group (Dr Viola, Dr Ganderton) worked in NN laboratories learning new methods and applying them to York-produced IR. All this contributed to setting of mutual trust and established common methodological and research platforms between leading insulin/health care industrial partner (NN) and York group. Collaborators from the IOCB (Prague) participated in some of these meetings. Possible involvement of this group in future, more expanded, work is considered. The recruitment to fill NN-sponsored PhD studentship, via GG Dodson Fund, was successful. The PhD student will perform IR-research that is complementary to the MRC Programme Grant. New collaboration with group of Prof P Jungwirth from IOCB, Prague, has been initiated. it will involve application of molecular simulations and dynamics to investigate conformational states of insulin in storage vesicles in pancreas. York will provide here crystal structures of differently liganded insulin, benefiting from novel insight into insulin-ligand interactions obtained via cutting-edge modelling simulations.
Collaborator Contribution see above - all are in confidential stage at this phase of the programme.
Impact All outputs are still in a confidential, pre-publication stage.
Start Year 2013
 
Description Main Collaborations within the MRC Programme grant 
Organisation University of York
Country United Kingdom 
Sector Academic/University 
PI Contribution The first year of this Programme involved collaborations between molecular/structural biology laboratory (York Structural Biology Laboratory (YSBL)- University of York), organic/protein chemistry (Institute of Organic Chemistry and Biochemistry Czech Academy of Sciences (IOCB) - Prague), cell biology (University College Cork (UCC)). The Walter and Eliza Hall Institute of Medical Research ((WEHI), Melbourne) deliver the know-how in large-scale of production of human insulin receptor (IR). All groups worked within the scope of the planned programme. YSBL work underpinned all other activities, focusing on development of sustainable and novel production of IR that is one of the major bottlenecks of insulin-related research. The Ysbl Eukarytioc Expression Facility (YEF) has been established in this process. It is self-sufficient for baculovirus and mammalian cloning and expression work for this project. Over 50 IR constructs have been generated, and expression of some is optimized. IOCB developed novel synthetic routes for generation of new insulin and IGF analogues. They also established binding and signaling essays for all IR isoforms, which serve the characterization and preliminary evaluation of the most promising analogues from steadily growing (>100) library of semi/fully-synthetic insulins generated in this project. More complex signaling networks have been studied further in the UCC. The first, large batch of IR for structural studies has also been provided by WEHI. Therefore the first year of this programme cemented its foundations and positively validated its organizational and scientific principles. The multifaceted character of this programme grant has already generated growing network of new collaborations. Notably, an agreement between the YSBL and Novo Nordisk A/S (Copenhagen) has been signed upon which a fund in memory of late Prof Guy Dodson has been established. £150K will be provided here by Novo-Nordisk towards the PhD studentships devoted to insulin-related research. The first PhD programme - closely linked with this MRC grant - will start in October 2014. This YSBL-Novo Nordisk PhD-based initiative generates an excellent and exciting opportunity to elevate the collaboration between these institutions into the field of diabetes research that will transgress existing limiting boundaries. Collaboration between Prof AM Brzozowski (YSBL) and Prof Pierre de Meyts (Professor Emeritus in the Dept. of Chemistry of Catholic University of Louvain (Belgium), ex-Director of Research of the Hagedorn Research Institute, Gentofte, DK)) has also been established in the field of insulin-related structural and cellular biology in insects. This aims to develop a new programme - tangential and complementary to this MRC award - that will be focused on insects' insulins (DLPs), IR and DLPs-binding proteins. This research - due to a remarkable similarity of human and insects' insulin-related biology - will provide an additional insight into specificity and structure-function relationships of insulins and IRs. It should also shed light on age-regulatory role of insulin and IGFs in humans. Several proteins have already been produced here and their complexes characterized by X-ray crystallography. Branching of this research into insects-based model of insulin function resulted also in a startup of collaboration between Prof AM Brzozowski (YSBL) and Dr Sean T Sweeney (Dept. of Biology, University of York). This should expand the studies on biology of insects' insulins/IR systems (initiated with Prof P de Meyts) into probing of the phenotype modulation by insulin-signaling related genes knock-outs/rescue-based studies. They may lead to insect-based models for diabetes and life-span regulation in humans. Dr Sweeney has already developed key tools for studies of synaptic transmission and growth in Drosophila that delivered important insight into some forms of human neurodegenerative diseases, and molecular bases of aging and pain. These molecular tools will be applied for insulin-related signaling and regulatory systems. The second year of this Programme Grant resulted in cementing of York collaboration with Novo Nordisk (Copenhagen)(NN). Series of meetings in York and Copenhagen yielded detail definition of the expansion of Insulin Receptor (IR) research programme that is the core of this award. The agreement about collaboration, including details of the intellectual properties, publication rights, patenting etc. has been signed by both sides. Most advanced methodology in IR characterisation (developed in Copenhagen) has been made available to York group. Members of the YSBL group (Dr Viola, Dr Ganderton) worked in NN laboratories learning new methods and applying them to York-produced IR. All this contributed to setting of mutual trust and established common methodological and research platforms between leading insulin/health care industrial partner (NN) and York group. Collaborators from the IOCB (Prague) participated in some of these meetings. Possible involvement of this group in future, more expanded, work is considered. The recruitment to fill NN-sponsored PhD studentship, via GG Dodson Fund, was successful. The PhD student will perform IR-research that is complementary to the MRC Programme Grant. New collaboration with group of Prof P Jungwirth from IOCB, Prague, has been initiated. it will involve application of molecular simulations and dynamics to investigate conformational states of insulin in storage vesicles in pancreas. York will provide here crystal structures of differently liganded insulin, benefiting from novel insight into insulin-ligand interactions obtained via cutting-edge modelling simulations.
Collaborator Contribution see above - all are in confidential stage at this phase of the programme.
Impact All outputs are still in a confidential, pre-publication stage.
Start Year 2013
 
Description Main Collaborations within the MRC Programme grant -Year 3 
Organisation Academy of Sciences of the Czech Republic
Department Institute of Organic Chemistry and Biochemistry
Country Czech Republic 
Sector Academic/University 
PI Contribution The first year of this Programme set up the collaborations between molecular/structural biology laboratory (York Structural Biology Laboratory (YSBL)- University of York), organic/protein chemistry (Institute of Organic Chemistry and Biochemistry Czech Academy of Sciences (IOCB) - Prague), cell biology (University College Cork (UCC)). The Walter and Eliza Hall Institute of Medical Research ((WEHI), Melbourne) deliver the know-how in large-scale of production of human insulin receptor (IR). All groups worked in year 3 within the scope of the planned programme. YSBL work underpinned all other activities, focusing on development of sustainable and novel production of IR that is one of the major bottlenecks of insulin-related research. The Ysbl Eukaryotic Expression Facility (YEF) that has been established in year 1 - to provide a self-sufficient platform for baculovirus and mammalian cloning and expression work for this project - was developed further. The second phase of the development of YEF was focused on assurance of high-capacity protein production. Over £20K pounds from University internal funds was invested into the additional cell culture-related equipment (shakers, bottle rollers, tangential flow media concentrators etc.) As the body of work in the protein production level has increased a new bottle neck: uninterrupted access to protein purification equipment has emerged. Therefore grant application for this project-devoted protein purification equipment has bee filed to Diabetes UK. It was successful, and the grant of a total £49K value was obtained. The equipment has been purchased and is instrumental for the success of this research. The complex issue of a most challenging production of IR constructs prompt Dr T Ganderton (one of the York Post-Docs in this project) to try development of an alternative mammalian-based systems for the expression of challenging proteins. He become a member of the BBSRC BioProNET network, and, subsequently applied within this initiative for a one-year grant "Hijacking intra-cellular storage bodies to create a novel mammalian cell-based expression system for the production of hard to express proteins". This application has been successful and one-year grant of total value £99,084 has been obtained. It should contribute to further progress of this project (starts 1.04.2016). The network of the existing collaborations has been cemented in the third year of the project. The already signed agreement between the YSBL and Novo Nordisk A/S (Copenhagen) delivered insulin-research oriented fund, in memory of late Prof Guy Dodson (GG Dodson Fund). £150K was assured here by Novo-Nordisk for the PhD studentships devoted to insulin-related research. The recruitment for the PhD studentship was successful, and it started October 2014. The programme of this PhD is very closely linked with the research of this MRC project. It allows exploration of new challenging lines of research, which would be difficult (time/personnel limitations) within the scope of this grant. The collaboration of York with the Novo Nordisk (NN) on Insulin Receptor (IR) has been successfully established. It provides our group a direct access to the unique NN know-how in functional characterisation of the IR. Post-docs employed in York on the MRC grant are fused into the network of personal collaborative contacts with NN, and had a working experience in its laboratories. The ongoing collaboration between Prof AM Brzozowski (YSBL) and Prof Pierre de Meyts (PDM, Professor Emeritus in the Dept. of Chemistry of Catholic University of Louvain (Belgium), ex-Director of Research of the Hagedorn Research Institute, Gentofte, DK)), and NN has been continued in the field of insulin-related structural and cellular biology in insects. Several insect insulin-binding proteins have been produced in free and complexes with insulin and similar hormones. Their crystal structures have also been obtained. They are subjects of an important, emerging manuscript of an alternative molecular regulation of insulin bio-availability. YSBL insulin group, PDM, NN and IOCB are working also on the development of a research programme that is tangential and complementary to this MRC award. It will be focused on insects' insulins (DLPs), IR and DLPs-binding proteins. This research - due to a remarkable conservation and similarity of human and insects' insulin-related biology - should provide a unique insight into specificity and structure-function relationships of insulins and IRs in the animal kingdom. The conservation of the insulin-axis in animals will impact our understanding of the role of insulin and IGFs in humans, not only in regulation of metabolism but in determination of life-span as well. New stream of research towards elucidation of impact of insulin-like hormones on Central Nervous System has also been initiated. The grant application is being written in collaboration with Dr Sean T Sweeney (Dept. of Biology, University of York), who has already developed key tools for studies of synaptic transmission and growth in Drosophila, which already delivered important insight into some forms of human neuro-degenerative diseases, and molecular bases of aging and pain. Series of meetings between York group, NN and IOCB provided outline of the expansion of Insulin Receptor (IR) research programme that is the core of this award. The discussion on a separate IOCB-NN-York agreement about insulin-focused collaboration is in progress. New collaboration with Computational Chemistry group of Prof P Jungwirth from IOCB, Prague, has also been established. It aimed to do molecular simulations and dynamics of conformational states of insulin in storage vesicles in pancreas. York/IOCB groups determined crystal structures of insulin with new, physiologically relevant, liganded insulin, feeding and correlating this data with Jungwirth results.
Collaborator Contribution A. The group of Dr Jiracek (IOCB) was/is instrumental for this research in the field of insulin. They: (i) semi-synthesised over 100 analogues of insulin, (ii) setup expression and production of IGF-1 and IGF-2 for insulin/IGFs hormone hybrid production, (iii) Produced insulin-IGF-1/IGF-2 hybrids to elucidate the role of IGFs domains in insulin/IGF-axis signalling, (iii) setup complex and very specific assays for insulin/analogues : IR/IGF1-R binding, receptor auto-phosphorylation, and cell-signalling (iv) started work towards determination of the physiological conformation of insulin in pancreas, (v) developed organic scaffolds for synthesis of organic mimics of insulin. B. The group of Dr Jungwirth has carried out independently molecular dynamic calculations and simulations of insulin in its physiological environment. C. Novo Nordisk provided on-going, and free access to their facilities, especially to assess the activity of IR constructs on their early expression stage and in their final pure form. D. York group was a member of MARIE SKLODOWSKA-CURIE ACTIONS: Innovative Training Networks (ITN) application that was co-ordinated by Prof R O'Connor from UCC. It grouped 9 academic centers and 5 industrial partners to work on Insulin/IGF system in health and disease. This application was not successful, but it will be improved and submitted again in 2016.
Impact (i) This collaborative (IOCB-York) research delivered the currently most 'metabolic' - highly IR-B isoform specific analogue of insulin. This type of analogues are highly desired to eliminate the unwanted side-effects of insulin: to streamline its glycemic control via 'metabolic' form of the IR (IR-B)(10.1038/srep19431). (ii) The 'metabolic' analogue of insulin is the subject of two patents has been filed: one Czech Rep. - approved: C73-14, one EU (final approval stage): PC1414PC00. (iii) The most-optimum organic scaffolds for organic mimics of insulin has been design, and synthesized (DOI: 10.3390/molecules201019310). (iv) The combinatorial libraries for synthesis of insulin mimics have been made, and first organic mimics of insulin have been synthesised (confidential - on-going work). (v) Novel, physiological allosteric regulator of insulin structure and aggregation have been identified by structural work and molecular simulations (manuscript in preparation). (vi) Two grants associated with this research has been secured (Diabetes UK (£49K), BBSRC BioProNET (£99K)) for the equipment and development of project-relevant novel expression systems. (vii) New molecular bases of the allosteric regulation of physiological states of insulin have been identified and characterised by structural and molecular dynamics studies (manuscript in preparation). (viii) First IR constructs have been obtained in York in quantities allowing initiation of structural studies, and crystals of one of the constructs have been obtained (confidential - ongoing work). This is a long awaited breakthrough in IR research. Till now, production of IR constructs for structural biology was possible only in the centers that could afford large (industrial) scale mammalian cell cultures. Here, we shown that this is possible on academic/regular laboratory level as well. This developments should have a most positive impact on an overall progress in IR and insulin research in general. (ix) The analysis of obtained crystal structures of Drosophila Impl2 insulin-binding protein complexes unraveled exciting relationships in insulin axis in the animal kingdom. These findings will have important impact on the understanding of human insulin signalling (manuscript in preparation). (x) The expertise of members of the York team in cloning and protein production of difficult proteins was tested on human heparanase (enzyme strongly linked with cancer metastasis), which was successfully produced, and its crystal structure has been solved in York as well (PubMed ID: 26575439).
Start Year 2013
 
Description Main Collaborations within the MRC Programme grant -Year 3 
Organisation Novo Nordisk
Country Denmark 
Sector Public 
PI Contribution The first year of this Programme set up the collaborations between molecular/structural biology laboratory (York Structural Biology Laboratory (YSBL)- University of York), organic/protein chemistry (Institute of Organic Chemistry and Biochemistry Czech Academy of Sciences (IOCB) - Prague), cell biology (University College Cork (UCC)). The Walter and Eliza Hall Institute of Medical Research ((WEHI), Melbourne) deliver the know-how in large-scale of production of human insulin receptor (IR). All groups worked in year 3 within the scope of the planned programme. YSBL work underpinned all other activities, focusing on development of sustainable and novel production of IR that is one of the major bottlenecks of insulin-related research. The Ysbl Eukaryotic Expression Facility (YEF) that has been established in year 1 - to provide a self-sufficient platform for baculovirus and mammalian cloning and expression work for this project - was developed further. The second phase of the development of YEF was focused on assurance of high-capacity protein production. Over £20K pounds from University internal funds was invested into the additional cell culture-related equipment (shakers, bottle rollers, tangential flow media concentrators etc.) As the body of work in the protein production level has increased a new bottle neck: uninterrupted access to protein purification equipment has emerged. Therefore grant application for this project-devoted protein purification equipment has bee filed to Diabetes UK. It was successful, and the grant of a total £49K value was obtained. The equipment has been purchased and is instrumental for the success of this research. The complex issue of a most challenging production of IR constructs prompt Dr T Ganderton (one of the York Post-Docs in this project) to try development of an alternative mammalian-based systems for the expression of challenging proteins. He become a member of the BBSRC BioProNET network, and, subsequently applied within this initiative for a one-year grant "Hijacking intra-cellular storage bodies to create a novel mammalian cell-based expression system for the production of hard to express proteins". This application has been successful and one-year grant of total value £99,084 has been obtained. It should contribute to further progress of this project (starts 1.04.2016). The network of the existing collaborations has been cemented in the third year of the project. The already signed agreement between the YSBL and Novo Nordisk A/S (Copenhagen) delivered insulin-research oriented fund, in memory of late Prof Guy Dodson (GG Dodson Fund). £150K was assured here by Novo-Nordisk for the PhD studentships devoted to insulin-related research. The recruitment for the PhD studentship was successful, and it started October 2014. The programme of this PhD is very closely linked with the research of this MRC project. It allows exploration of new challenging lines of research, which would be difficult (time/personnel limitations) within the scope of this grant. The collaboration of York with the Novo Nordisk (NN) on Insulin Receptor (IR) has been successfully established. It provides our group a direct access to the unique NN know-how in functional characterisation of the IR. Post-docs employed in York on the MRC grant are fused into the network of personal collaborative contacts with NN, and had a working experience in its laboratories. The ongoing collaboration between Prof AM Brzozowski (YSBL) and Prof Pierre de Meyts (PDM, Professor Emeritus in the Dept. of Chemistry of Catholic University of Louvain (Belgium), ex-Director of Research of the Hagedorn Research Institute, Gentofte, DK)), and NN has been continued in the field of insulin-related structural and cellular biology in insects. Several insect insulin-binding proteins have been produced in free and complexes with insulin and similar hormones. Their crystal structures have also been obtained. They are subjects of an important, emerging manuscript of an alternative molecular regulation of insulin bio-availability. YSBL insulin group, PDM, NN and IOCB are working also on the development of a research programme that is tangential and complementary to this MRC award. It will be focused on insects' insulins (DLPs), IR and DLPs-binding proteins. This research - due to a remarkable conservation and similarity of human and insects' insulin-related biology - should provide a unique insight into specificity and structure-function relationships of insulins and IRs in the animal kingdom. The conservation of the insulin-axis in animals will impact our understanding of the role of insulin and IGFs in humans, not only in regulation of metabolism but in determination of life-span as well. New stream of research towards elucidation of impact of insulin-like hormones on Central Nervous System has also been initiated. The grant application is being written in collaboration with Dr Sean T Sweeney (Dept. of Biology, University of York), who has already developed key tools for studies of synaptic transmission and growth in Drosophila, which already delivered important insight into some forms of human neuro-degenerative diseases, and molecular bases of aging and pain. Series of meetings between York group, NN and IOCB provided outline of the expansion of Insulin Receptor (IR) research programme that is the core of this award. The discussion on a separate IOCB-NN-York agreement about insulin-focused collaboration is in progress. New collaboration with Computational Chemistry group of Prof P Jungwirth from IOCB, Prague, has also been established. It aimed to do molecular simulations and dynamics of conformational states of insulin in storage vesicles in pancreas. York/IOCB groups determined crystal structures of insulin with new, physiologically relevant, liganded insulin, feeding and correlating this data with Jungwirth results.
Collaborator Contribution A. The group of Dr Jiracek (IOCB) was/is instrumental for this research in the field of insulin. They: (i) semi-synthesised over 100 analogues of insulin, (ii) setup expression and production of IGF-1 and IGF-2 for insulin/IGFs hormone hybrid production, (iii) Produced insulin-IGF-1/IGF-2 hybrids to elucidate the role of IGFs domains in insulin/IGF-axis signalling, (iii) setup complex and very specific assays for insulin/analogues : IR/IGF1-R binding, receptor auto-phosphorylation, and cell-signalling (iv) started work towards determination of the physiological conformation of insulin in pancreas, (v) developed organic scaffolds for synthesis of organic mimics of insulin. B. The group of Dr Jungwirth has carried out independently molecular dynamic calculations and simulations of insulin in its physiological environment. C. Novo Nordisk provided on-going, and free access to their facilities, especially to assess the activity of IR constructs on their early expression stage and in their final pure form. D. York group was a member of MARIE SKLODOWSKA-CURIE ACTIONS: Innovative Training Networks (ITN) application that was co-ordinated by Prof R O'Connor from UCC. It grouped 9 academic centers and 5 industrial partners to work on Insulin/IGF system in health and disease. This application was not successful, but it will be improved and submitted again in 2016.
Impact (i) This collaborative (IOCB-York) research delivered the currently most 'metabolic' - highly IR-B isoform specific analogue of insulin. This type of analogues are highly desired to eliminate the unwanted side-effects of insulin: to streamline its glycemic control via 'metabolic' form of the IR (IR-B)(10.1038/srep19431). (ii) The 'metabolic' analogue of insulin is the subject of two patents has been filed: one Czech Rep. - approved: C73-14, one EU (final approval stage): PC1414PC00. (iii) The most-optimum organic scaffolds for organic mimics of insulin has been design, and synthesized (DOI: 10.3390/molecules201019310). (iv) The combinatorial libraries for synthesis of insulin mimics have been made, and first organic mimics of insulin have been synthesised (confidential - on-going work). (v) Novel, physiological allosteric regulator of insulin structure and aggregation have been identified by structural work and molecular simulations (manuscript in preparation). (vi) Two grants associated with this research has been secured (Diabetes UK (£49K), BBSRC BioProNET (£99K)) for the equipment and development of project-relevant novel expression systems. (vii) New molecular bases of the allosteric regulation of physiological states of insulin have been identified and characterised by structural and molecular dynamics studies (manuscript in preparation). (viii) First IR constructs have been obtained in York in quantities allowing initiation of structural studies, and crystals of one of the constructs have been obtained (confidential - ongoing work). This is a long awaited breakthrough in IR research. Till now, production of IR constructs for structural biology was possible only in the centers that could afford large (industrial) scale mammalian cell cultures. Here, we shown that this is possible on academic/regular laboratory level as well. This developments should have a most positive impact on an overall progress in IR and insulin research in general. (ix) The analysis of obtained crystal structures of Drosophila Impl2 insulin-binding protein complexes unraveled exciting relationships in insulin axis in the animal kingdom. These findings will have important impact on the understanding of human insulin signalling (manuscript in preparation). (x) The expertise of members of the York team in cloning and protein production of difficult proteins was tested on human heparanase (enzyme strongly linked with cancer metastasis), which was successfully produced, and its crystal structure has been solved in York as well (PubMed ID: 26575439).
Start Year 2013
 
Description Main Collaborations within the MRC Programme grant -Year 3 
Organisation University College Cork
Country Ireland 
Sector Academic/University 
PI Contribution The first year of this Programme set up the collaborations between molecular/structural biology laboratory (York Structural Biology Laboratory (YSBL)- University of York), organic/protein chemistry (Institute of Organic Chemistry and Biochemistry Czech Academy of Sciences (IOCB) - Prague), cell biology (University College Cork (UCC)). The Walter and Eliza Hall Institute of Medical Research ((WEHI), Melbourne) deliver the know-how in large-scale of production of human insulin receptor (IR). All groups worked in year 3 within the scope of the planned programme. YSBL work underpinned all other activities, focusing on development of sustainable and novel production of IR that is one of the major bottlenecks of insulin-related research. The Ysbl Eukaryotic Expression Facility (YEF) that has been established in year 1 - to provide a self-sufficient platform for baculovirus and mammalian cloning and expression work for this project - was developed further. The second phase of the development of YEF was focused on assurance of high-capacity protein production. Over £20K pounds from University internal funds was invested into the additional cell culture-related equipment (shakers, bottle rollers, tangential flow media concentrators etc.) As the body of work in the protein production level has increased a new bottle neck: uninterrupted access to protein purification equipment has emerged. Therefore grant application for this project-devoted protein purification equipment has bee filed to Diabetes UK. It was successful, and the grant of a total £49K value was obtained. The equipment has been purchased and is instrumental for the success of this research. The complex issue of a most challenging production of IR constructs prompt Dr T Ganderton (one of the York Post-Docs in this project) to try development of an alternative mammalian-based systems for the expression of challenging proteins. He become a member of the BBSRC BioProNET network, and, subsequently applied within this initiative for a one-year grant "Hijacking intra-cellular storage bodies to create a novel mammalian cell-based expression system for the production of hard to express proteins". This application has been successful and one-year grant of total value £99,084 has been obtained. It should contribute to further progress of this project (starts 1.04.2016). The network of the existing collaborations has been cemented in the third year of the project. The already signed agreement between the YSBL and Novo Nordisk A/S (Copenhagen) delivered insulin-research oriented fund, in memory of late Prof Guy Dodson (GG Dodson Fund). £150K was assured here by Novo-Nordisk for the PhD studentships devoted to insulin-related research. The recruitment for the PhD studentship was successful, and it started October 2014. The programme of this PhD is very closely linked with the research of this MRC project. It allows exploration of new challenging lines of research, which would be difficult (time/personnel limitations) within the scope of this grant. The collaboration of York with the Novo Nordisk (NN) on Insulin Receptor (IR) has been successfully established. It provides our group a direct access to the unique NN know-how in functional characterisation of the IR. Post-docs employed in York on the MRC grant are fused into the network of personal collaborative contacts with NN, and had a working experience in its laboratories. The ongoing collaboration between Prof AM Brzozowski (YSBL) and Prof Pierre de Meyts (PDM, Professor Emeritus in the Dept. of Chemistry of Catholic University of Louvain (Belgium), ex-Director of Research of the Hagedorn Research Institute, Gentofte, DK)), and NN has been continued in the field of insulin-related structural and cellular biology in insects. Several insect insulin-binding proteins have been produced in free and complexes with insulin and similar hormones. Their crystal structures have also been obtained. They are subjects of an important, emerging manuscript of an alternative molecular regulation of insulin bio-availability. YSBL insulin group, PDM, NN and IOCB are working also on the development of a research programme that is tangential and complementary to this MRC award. It will be focused on insects' insulins (DLPs), IR and DLPs-binding proteins. This research - due to a remarkable conservation and similarity of human and insects' insulin-related biology - should provide a unique insight into specificity and structure-function relationships of insulins and IRs in the animal kingdom. The conservation of the insulin-axis in animals will impact our understanding of the role of insulin and IGFs in humans, not only in regulation of metabolism but in determination of life-span as well. New stream of research towards elucidation of impact of insulin-like hormones on Central Nervous System has also been initiated. The grant application is being written in collaboration with Dr Sean T Sweeney (Dept. of Biology, University of York), who has already developed key tools for studies of synaptic transmission and growth in Drosophila, which already delivered important insight into some forms of human neuro-degenerative diseases, and molecular bases of aging and pain. Series of meetings between York group, NN and IOCB provided outline of the expansion of Insulin Receptor (IR) research programme that is the core of this award. The discussion on a separate IOCB-NN-York agreement about insulin-focused collaboration is in progress. New collaboration with Computational Chemistry group of Prof P Jungwirth from IOCB, Prague, has also been established. It aimed to do molecular simulations and dynamics of conformational states of insulin in storage vesicles in pancreas. York/IOCB groups determined crystal structures of insulin with new, physiologically relevant, liganded insulin, feeding and correlating this data with Jungwirth results.
Collaborator Contribution A. The group of Dr Jiracek (IOCB) was/is instrumental for this research in the field of insulin. They: (i) semi-synthesised over 100 analogues of insulin, (ii) setup expression and production of IGF-1 and IGF-2 for insulin/IGFs hormone hybrid production, (iii) Produced insulin-IGF-1/IGF-2 hybrids to elucidate the role of IGFs domains in insulin/IGF-axis signalling, (iii) setup complex and very specific assays for insulin/analogues : IR/IGF1-R binding, receptor auto-phosphorylation, and cell-signalling (iv) started work towards determination of the physiological conformation of insulin in pancreas, (v) developed organic scaffolds for synthesis of organic mimics of insulin. B. The group of Dr Jungwirth has carried out independently molecular dynamic calculations and simulations of insulin in its physiological environment. C. Novo Nordisk provided on-going, and free access to their facilities, especially to assess the activity of IR constructs on their early expression stage and in their final pure form. D. York group was a member of MARIE SKLODOWSKA-CURIE ACTIONS: Innovative Training Networks (ITN) application that was co-ordinated by Prof R O'Connor from UCC. It grouped 9 academic centers and 5 industrial partners to work on Insulin/IGF system in health and disease. This application was not successful, but it will be improved and submitted again in 2016.
Impact (i) This collaborative (IOCB-York) research delivered the currently most 'metabolic' - highly IR-B isoform specific analogue of insulin. This type of analogues are highly desired to eliminate the unwanted side-effects of insulin: to streamline its glycemic control via 'metabolic' form of the IR (IR-B)(10.1038/srep19431). (ii) The 'metabolic' analogue of insulin is the subject of two patents has been filed: one Czech Rep. - approved: C73-14, one EU (final approval stage): PC1414PC00. (iii) The most-optimum organic scaffolds for organic mimics of insulin has been design, and synthesized (DOI: 10.3390/molecules201019310). (iv) The combinatorial libraries for synthesis of insulin mimics have been made, and first organic mimics of insulin have been synthesised (confidential - on-going work). (v) Novel, physiological allosteric regulator of insulin structure and aggregation have been identified by structural work and molecular simulations (manuscript in preparation). (vi) Two grants associated with this research has been secured (Diabetes UK (£49K), BBSRC BioProNET (£99K)) for the equipment and development of project-relevant novel expression systems. (vii) New molecular bases of the allosteric regulation of physiological states of insulin have been identified and characterised by structural and molecular dynamics studies (manuscript in preparation). (viii) First IR constructs have been obtained in York in quantities allowing initiation of structural studies, and crystals of one of the constructs have been obtained (confidential - ongoing work). This is a long awaited breakthrough in IR research. Till now, production of IR constructs for structural biology was possible only in the centers that could afford large (industrial) scale mammalian cell cultures. Here, we shown that this is possible on academic/regular laboratory level as well. This developments should have a most positive impact on an overall progress in IR and insulin research in general. (ix) The analysis of obtained crystal structures of Drosophila Impl2 insulin-binding protein complexes unraveled exciting relationships in insulin axis in the animal kingdom. These findings will have important impact on the understanding of human insulin signalling (manuscript in preparation). (x) The expertise of members of the York team in cloning and protein production of difficult proteins was tested on human heparanase (enzyme strongly linked with cancer metastasis), which was successfully produced, and its crystal structure has been solved in York as well (PubMed ID: 26575439).
Start Year 2013
 
Title Insulin derivative of cyclic structure in B-chain C-terminus 
Description Modification of the B-chain of the C-terminus that lead to the semi-synthesis of highly metabolic insulin analogue: i.e. a hormone that has a very high affinity for the metabolic isoform (IR-B) of human Insulin Receptor. 
IP Reference Czech Republic Patent - 2014-450 
Protection Patent granted
Year Protection Granted 2016
Licensed No
Impact Not yet.
 
Title Insulin derivative with cyclic structure in the C-terminus of the B-chain 
Description The is the EU Patent application. It describes chemical modification of human insulin C-terminal part of the B-chain of this hormone. This modification gives this insulin analogue almost 3 times higher preference for the isoform B of the insulin receptor (IR-B). This one of the two isoforms of IR that is mainly responsible for fully metabolic and glycemic responses to insulin signalling. The other IR isoform - IR-A - is a good binder of Insulin-like Growth Factors 1 and 2, hence can stimulate unwanted mitogenic responses through the IR. As IR-B forms more than 90% of IR in liver - the main glycemic response organ - it is important that novel insulin analogues will have a higher affinity to IR-B, to mimic better physiological profile of insulin. The analogue described in this application shows directions in designing of novel types of hormones with such desired metabolic properties. 
IP Reference 15745135.2-1402 
Protection Patent application published
Year Protection Granted
Licensed No
Impact The work here is in progress.
 
Title Insulin derivative with cyclic structure in the C-terminus of the B-chain 
Description This invention describes a new derivative of human insulin (referred here as to derivative I) which has a cyclic structure in the C-terminus of the B-chain. The side chains of B26 and B29 are replaced - and crosslinked - by a triazole bridge which enforces specific conformation of insulin B23-B30 chain. The derivative I exhibits a higher affinity to both isoforms of the insulin receptor than that of human insulin, and at the same time, it has a markedly higher affinity to the predominantly metabolic B isoform than to the predominantly mitogenic A isoform of this receptor. Another advantage is a significantly lower affinity of the derivative to IGF-1R than that of human insulin. These properties predetermine the new insulin derivative of formula I for use as effectively acting insulin, lowering blood glucose levels in vivo, with lower side effects, in particular, a lower possibility of developing cancer, than those of human insulin. Insulin derivative I exhibits two-fold higher binding affinity for the insulin receptor (IR-A isoform) in the membranes of human IM-9 lymphocytes and particularly five-fold higher binding affinity for the insulin receptor in the membranes of murine fibroblasts transfected with the B isoform (IR-B) of the human insulin receptor that of human insulin. Hence, the derivative is 2-5-fold more active more towards insulin receptor than human insulin and moreover with a significant preference for the metabolic isoform B of IR. Besides this, the derivative of formula I displays 5-fold lower binding affinity than human insulin against the IGF-1 receptor (IGF-1R)) in the membranes of mouse fibroblasts transfected with human IGF-1R. Thus, the novel insulin derivative I combines in its properties not only the preferential affinity for IR-B, which is the metabolic isoform of insulin receptor, but also very high binding affinity generally (515% compared to human insulin) and, hence, insulin derivative I belongs to the most potent insulin derivatives ever. Especially, the low affinity of the derivative of formula I towards IGF-1 receptor is unique. Similar properties have not been described yet for any other insulin derivative and predetermine the derivative of formula I for the clinical use to effectively lower the blood glucose levels in diabetics with a lower risk of the development of cancer (due to the low binding to IGF-1R). The almost 5-fold higher binding affinity than that of human insulin for IR-B isoform may also enable the use of lower doses of this derivative than of human insulin, which could result in a cheaper treatment. 
IP Reference EP3160993B1 
Protection Patent granted
Year Protection Granted 2018
Licensed No
Impact The invention relates to a new insulin derivative with high binding affinity to both isoforms of the insulin receptor. Affinity to isoform B is significantly higher than affinity to isoform A; the derivative also exhibits a very low affinity to the IGF-1 receptor. The insulin derivative patented here, according to this invention, is suitable as an active ingredient in pharmaceutical compositions for treating or preventing diabetes and conditions characterized by elevated levels of blood glucose, i.e. for the reduction of blood glucose concentration. Due to its high affinity for metabolic isoform B of insulin receptor, and very low affinity for the receptor for IGF-1 and reduced ability to dimerize, the derivative has a significant potential for use as a preparation with a rapid onset of action and a reduced risk of developing cancer. This invention opened new directions for design and production of insulin analogues with high, desired affinities for the metabolic form of the insulin receptor.
 
Title Insulin derivative with cyclic structure in the C-terminus of the B-chain 
Description This patent describe design, synthesis and evaluation of human insulin analogues with application of click chemistry to a rational steering of hormone's structure and functionality. Promising analogues with affinities towards specific isoforms of human Insulin Receptor have been obtained. Continuation of this work may result in insulin analogues with full 'metabolic' profile/signalling properties. 
IP Reference WO2016000667 
Protection Patent granted
Year Protection Granted 2013
Licensed No
Impact This results of this research proved that it is possible to achieve novel insulin analogues with desired properties. Insulin analogues with high affinities towards IR-B - metabolic form of insulin receptor - are of a wide clinical applications. This work is an important step towards this direction. It opened new stream of our research on insulin (and IGF) analogues showing also potential of application of organic chemistry in rational modification of insulin.
 
Description Charity Fundrising: event Fundraising for Diabetes UK 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach Regional
Primary Audience Public/other audiences
Results and Impact To happen: 2 March 2016, Ripley's Castle
Year(s) Of Engagement Activity 2016
 
Description European Researchers Night: Yornight 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? Yes
Geographic Reach Local
Primary Audience Public/other audiences
Results and Impact A group of researchers from this MRC Programme Grant hold an open exhibition/stand at York Medical Society building in York. It was open to the public 4-9 pm on the 26th of September. The role of insulin in health and disease was presented together with key aspects of our research. It was also linked with 'live' demonstration of X-ray crystallography based methodology and its impact on biomedical research. Beside local, York citizens, the exhibition/stand was visited by many foreign tourist. This provided an additional impact to this activity. Overall, the main importance of this activity was in expanding of public awareness about complexity of biomedical research, and some common molecular patterns in complex human disease such diabetes and cancer.

A very advanced biomedical research was presented to the public, highlighting also its social and economical importance. Inquiries of the members of the public covered a wide range of aspects. They concerned molecular complexity of diabetes and cancer, challenges for biomedical research in this field, main financial obstacles, alternative approaches to alleviate type 2 diabetes epidemics.
Year(s) Of Engagement Activity 2014
URL http://yornight.com/activities/
 
Description Public Outreach: Networking Day for Diabetes UK Local Support Groups 
Form Of Engagement Activity A formal working group, expert panel or dialogue
Part Of Official Scheme? No
Geographic Reach Regional
Primary Audience Patients, carers and/or patient groups
Results and Impact Circa 50 Diabetes UK volunteers of Local (North Yorkshire) Support Groups attended yearly meeting. They were informed by AM Brzozowski about the diabetes-related research in York and the main problems in insulin-related research. The lively discussion that followed underlined the need for such talks: importance of informing disease-linked volunteers about the progress and challenges of the research relevant to the pathology they are helping with.
Year(s) Of Engagement Activity 2015
URL https://www.diabetes.org.uk/How_we_help/Care-events/Volunteering/