Creating an effective and non toxic blood substitute
Lead Research Organisation:
University of Essex
Department Name: Biological Sciences
Abstract
Blood transfusion is a life saving technology. However red blood cells: have a limited shelf life; need blood group typing; cannot be used in immune compromised individuals; are not available at the scenes of accidents, battlefields or major emergencies; and are susceptible to viral contamination. Blood transfusion is big business. In the UK alone two million units of blood are transfused annually at a cost of > £200 million. The addressable future global market value is $10Bn. A number of companies have attempted to develop alternatives to transfusions based on modifications of the red cell oxygen transport protein hemoglobin. However, toxic side effects have been seen in clinical trials. Arising out of BBSRC funded grants, the University of Essex has engineered novel hemoglobin molecules designed to reduce cell damage. A patent is pending. The purpose of this grant is to undertake lead optimisation, product development and in vivo testing on this next generation blood substitute.
Organisations
Publications
Alomari E
(2018)
High- and low-affinity PEGylated hemoglobin-based oxygen carriers: Differential oxidative stress in a Guinea pig transfusion model.
in Free radical biology & medicine
Cooper
(2016)
Blood: A Very Short Introduction
Cooper CE
(2020)
Engineering hemoglobin to enable homogenous PEGylation without modifying protein functionality.
in Biomaterials science
Cooper CE
(2021)
Stability of Maleimide-PEG and Mono-Sulfone-PEG Conjugation to a Novel Engineered Cysteine in the Human Hemoglobin Alpha Subunit.
in Frontiers in chemistry
Cooper CE
(2019)
Engineering tyrosine residues into hemoglobin enhances heme reduction, decreases oxidative stress and increases vascular retention of a hemoglobin based blood substitute.
in Free radical biology & medicine
Rong Z
(2016)
Hemoglobin Effects on Nitric Oxide Mediated Hypoxic Vasodilation.
in Advances in experimental medicine and biology
Silaghi-Dumitrescu R
(2014)
Nitrite binding to globins: linkage isomerism, EPR silence and reductive chemistry.
in Nitric oxide : biology and chemistry
Silaghi-Dumitrescu R
(2015)
The reaction of oxyhemoglobin with nitric oxide: EPR evidence for an iron(III)-nitrate intermediate
in Inorganica Chimica Acta
Silkstone GG
(2016)
Engineering tyrosine electron transfer pathways decreases oxidative toxicity in hemoglobin: implications for blood substitute design.
in The Biochemical journal
Silkstone GGA
(2018)
Novel Redox Active Tyrosine Mutations Enhance the Regeneration of Functional Oxyhemoglobin from Methemoglobin: Implications for Design of Blood Substitutes.
in Advances in experimental medicine and biology
| Description | The project has enabled us to optimise our through protein electron transfer pathways in human hemoglobin (Hb) with the specific aim of developing a product for preclinical animal testing. The aim was to enhance the ferryl heme iron reduction in the presence of plasma reductants, thus weakening the potential for redox-induced oxidative damage. Our data demonstrated (1) the addition of protein electron pathways enhances ferryl reduction kinetics by plasma level reductants compared to wild type protein, decreasing oxidation of lipid membranes. (2) The position of the mutation to introduce this pathway greatly affected protein stability and oxygen binding cooperativity (3). The position of the mutation in the lead compound (ß-T84Y), in addition to enhanced ferryl reduction, also introduced a hitherto unseen advantage of enhanced ferric reductase activity (4). The mutation led to significant reduction in the capacity of Hb to induce pro-oxidant reactions (5). PEGylation (conjugation of Hb with polyethylene glycol) using the Euro-PEG method (via lysine residues) did not prevent the enhanced ferryl / ferric reduction kinetics, but did lead to increased oxygen affinity and decreased cooperativity. The adverse effects on O2 affinity and cooperativity induced by PEGylation appear inherent to nonspecific PEGylation methods, e.g. Sangart's clinically tested MP4OX showed an even more extreme effect than EUROPEGHb. However, an additional objective of the SFoF was to develop and test bespoke engineered -SH reactive sites for PEGylation. These have proven very successful, enabling homogeneous PEGylation by conjugation with a single PEGylation per aß dimer. We have achieved this with two linkers; Maleimide-PEG (20 kDa) and a novel CyPEG linker created by our collaborators Abzena. Both systems are equally homogeneous, however, CyPEG has the additional advantage of extra stability, degrading more slowly that the Mal-PEG conjugated protein. Equivalent techniques such as used on the EUROPEG-Hb system shows heterogeneous addition of PEG (5.6 kDa) from 1 to 8 sites per dimer. Our novel PEGylation novel system has no adverse effects on the affinity or cooperativity of oxygen binding. To our knowledge this is an exclusive property of our system. |
| Exploitation Route | The value endpoint for our product will be after phase I/II clinical trials. We perceive two possible routes to achieve this goal: Our favoured route would be to seek funding through an MRC DPFS pathway. Inneed we have recently been award a further DPFS award entitled "Optimisation of the manufacture of a homogeneous synthetic haemoglobin as a novel Oxygen Therapeutic / Blood Substitute (Ref MR/T025441/1)". this project is due to start in June 2020. This project will focus on two aspects. (1) To develop a manufacturing pathway to clinical trials for our product, generating a scalable GMP process will be developed in collaboration with CPI, a high value manufacturing catapult. (2) A highly focused set of pre-clinical toxicological testing and haemorrhagic shock model to show that we have overcome the issues of previous generations of HBOCs and to demonstrate the efficacy of the product with the homogenous PEGylation. Future funding avenues being explored for commercialisation include Innovate UK and continuation of the MRC DPFS. A recently awarded Enterprise Research Project Fund will examine potential markets and generate a techno-economic model with the aim of steering future commercialisation routes and attracting development partners. Early talks with potential investors and funding partners have given a strong steer towards use of the product in organ preservation for transplantation, which can generate clinical impact. |
| Sectors | Healthcare,Pharmaceuticals and Medical Biotechnology |
| Description | Manufacturing and commercial development of an arterial blood substitute/oxygen therapeutic |
| Amount | £33,036 (GBP) |
| Funding ID | 13207 |
| Organisation | University of Essex |
| Sector | Academic/University |
| Country | United Kingdom |
| Start | 03/2020 |
| End | 04/2020 |
| Description | Optimisation of the manufacture of a homogeneous synthetic haemoglobin as a novel Oxygen Therapeutic / Blood Substitute |
| Amount | £841,264 (GBP) |
| Funding ID | MR/T025441/1 |
| Organisation | Medical Research Council (MRC) |
| Sector | Public |
| Country | United Kingdom |
| Start | 11/2020 |
| End | 10/2022 |
| Title | MODIFIED GLOBIN PROTEINS |
| Description | Certain aspects of the present invention relate to modified proteins e.g. oxygen-carrying proteins, comprising at least one modification for homogenous conjugation of one or more polymeric moieties e.g. polyethylene glycol or derivatives thereof. |
| IP Reference | WO2019122871 |
| Protection | Patent application published |
| Year Protection Granted | 2019 |
| Licensed | No |
| Impact | No Im,pact as yet, used to protect IP and gain funding towards manufacture and clinical trials. |
| Title | MODIFIED GLOBIN PROTEINS WITH ALTERED ELECTRON TRANSPORT PATHWAY |
| Description | The present invention relates to a modified porphyrin-based oxygen-carrying protein, such as haemoglobin, which has been found, in its unmodified state to have a low affinity site of electron transfer and a high affinity electron transfer between a reductant and ferryl haem iron via one or more protein amino acids. The invention provides such proteins that comprise a modification to enhance this pathway. |
| IP Reference | WO2009004309 |
| Protection | Patent granted |
| Year Protection Granted | 2009 |
| Licensed | No |
| Impact | None |
| Title | MODIFIED HAEMOGLOBIN PROTEINS |
| Description | The present invention relates to modified proteins e.g. oxygen-carrying proteins, with improved or enhanced, in comparison to a reference protein, reduction of a metal ion associated with the modified protein. The present invention also relates to methods of using such modified proteins and compositions comprising such proteins e.g. in therapy. |
| IP Reference | WO2018167469 |
| Protection | Patent application published |
| Year Protection Granted | 2018 |
| Licensed | No |
| Impact | No impact as yet. Patent aplications used to protect IP and to forward research toward clinical trials. |
| Company Name | Cymblood |
| Description | Currently a shell company primed to accept funding for phase I trials. |
| Year Established | 2015 |
| Impact | None as yet. |
| Website | http://www.cymblood.com |
| Description | Article about the BBSRC and MRC funded research in the University of Essex "Essex Effect" magazine, aimed at alumni and University supporters. |
| Form Of Engagement Activity | A magazine, newsletter or online publication |
| Part Of Official Scheme? | No |
| Geographic Reach | Local |
| Primary Audience | Other audiences |
| Results and Impact | An annual magazine aimed at university alumni and supporters, this magazine is send to over 50,000 alumni worldwide - and many more read it online. |
| Year(s) Of Engagement Activity | 2013,2014,2015,2016,2017,2018 |
| URL | https://issuu.com/uniofessex/docs/ee-2018-singles-digital-online-v1 |
| Description | Conference presentation "Artificial Cells, Blood Substitutes & Nanomedicine" McGill University, 13-15 November, 2017, Montreal, Quebec, Canada |
| Form Of Engagement Activity | A talk or presentation |
| Part Of Official Scheme? | No |
| Geographic Reach | International |
| Primary Audience | Study participants or study members |
| Results and Impact | Conference presentation "Artificial Cells, Blood Substitutes & Nanomedicine" McGill University, 13-15 November, 2017, Montreal, Quebec, Canada |
| Year(s) Of Engagement Activity | 2017 |
| URL | http://www.medicine.mcgill.ca/artcell/ |
| Description | Knowledge Transfer Partnership (KTP) Research Showcase 2021 |
| Form Of Engagement Activity | Participation in an activity, workshop or similar |
| Part Of Official Scheme? | No |
| Geographic Reach | National |
| Primary Audience | Industry/Business |
| Results and Impact | The Knowledge Transfer Partnership (KTP) Research Showcase 2021research to other researchers, businesses and Innovate UK. |
| Year(s) Of Engagement Activity | 2021 |
| URL | https://www.essex.ac.uk/news/2021/10/21/essex-ktp-awards-2021 |
| Description | Press Prelease |
| Form Of Engagement Activity | A magazine, newsletter or online publication |
| Part Of Official Scheme? | No |
| Geographic Reach | Regional |
| Primary Audience | Media (as a channel to the public) |
| Results and Impact | Press release of funding news for the latest stage of Blood Substitute/Oxygen Tharpeutics development. This went out to the University of Essex and local meadia via the University of Essex Communications Office. |
| Year(s) Of Engagement Activity | 2021 |
| URL | https://www.essex.ac.uk/news/2021/01/15/blood-substitute-research-enters-exciting-new-phase |
| Description | Public Engagement |
| Form Of Engagement Activity | Participation in an activity, workshop or similar |
| Part Of Official Scheme? | No |
| Geographic Reach | Local |
| Primary Audience | Public/other audiences |
| Results and Impact | Pint of Science is an annual science festival that aims to communicate contemporary scientific developments to the public by bringing scientists to pubs, cafés and other public places to share their research and findings. This was attended by ~60 members of the public at the "Other Monkey Brewing", 5-6 St Nicholas St, Colchester CO1 1DN on 9th May 2022. There were several talks designed to convey scientific research to the public. My contribution as entitled "A pint of artificial blood: Creating a safe blood substitute for the 21st century". |
| Year(s) Of Engagement Activity | 2022 |
| URL | https://pintofscience.co.uk/event/made-to-order-science |
| Description | Scienctific conference: International Society on Oxygen Transport to Tissue (ISOTT) |
| Form Of Engagement Activity | A talk or presentation |
| Part Of Official Scheme? | No |
| Geographic Reach | International |
| Primary Audience | Professional Practitioners |
| Results and Impact | Presented and discussed research at the conference of the International Society on Oxygen Transport to Tissue (ISOTT,.18-23rd September 2022, Ascona, Switzerland. Research talk entitled "A multi-targeted approach to engineer a safer generation of hemoglobin-based oxygen carrier". |
| Year(s) Of Engagement Activity | 2022 |
| URL | https://isott2022.com/ |
| Description | Talk at the XVII ISBS International Symposium Blood Substitutes and Oxygen Therapeutics, 21st-24th November 2019, Nara, Japan. |
| Form Of Engagement Activity | A talk or presentation |
| Part Of Official Scheme? | No |
| Geographic Reach | International |
| Primary Audience | Professional Practitioners |
| Results and Impact | Gave a talk on the deveplment of blood substitutes / oxygen therapeutics entitled "A uniquely homogenous PEGylation method without significantly affecting hemoglobin oxygen affinity and cooperativity" . |
| Year(s) Of Engagement Activity | 2019 |
| URL | http://www.pac-mice.jp/isbs2019/program.html |
| Description | University open day presentation on the creation of artificial blood substitutes. |
| Form Of Engagement Activity | A talk or presentation |
| Part Of Official Scheme? | No |
| Geographic Reach | National |
| Primary Audience | Public/other audiences |
| Results and Impact | University open day presentation on the creation of artificial blood substitutes. The open days are for public, prospective students and parents. These are help typically 7-10 times every year and include talks and demonstrations. |
| Year(s) Of Engagement Activity | 2016,2017,2018,2019 |
| URL | https://www.essex.ac.uk/visit-us/open-days |