From a Discovery to Clinical Applications Worldwide

A team from the University of Warsaw discovered universal analogous endings for mRNA molecules (so called 5’ caps) which guarantee higher mRNA stability in cell natural environment and enhance the production of desired proteins.

A discovery gives new hope to millions of people suffering from cancers, inborn genetic defects and to patients awaiting autotransplantation or regeneration of damaged or missing tissues. This is a new chapter in the development of effective genetic vaccines which, in the near future, will make it possible to fight malignancies, among others.

Effective commercialization of discoveriesresearch & development results

Enhancing mRNA Stability

University Technology Transfer Centre

www.uott.uw.edu.pl

The scientific world have concluded that in order to induce certain effects in gene therapies it is better to improve methods for specific mRNA molecule synthesis (several times less complex compared to DNA molecules) rather than interfere in DNA itself.

In contrast to DNA, mRNA is outside the cell nucleus, hence it is easier to introduce it to the cell as there is no need to interfere in the nucleus. Various mRNA molecules present in cells serve as “recipes” for production of particular proteins. These are

instructions based on which ribosomes, by decoding the information there contained, produce a particular, specific protein. Together with the rapid development in genetics, the interests of scientists and pharmaceutical companies have focused on studying the role of mRNA in treating diseases which result either from the overproduction of certain proteins (e.g., in cancer cell formation processes), or from chronic deficiency of some proteins in the body. In the scientific world, there is a belief that the ability to “manage” the production of particular proteins in practice will enable the development of highly effective therapies, including these against cancers.

mRNA is a molecule of ribonucleic acid (RNA) which functions as a messenger. Its role is to transfer genetic information from the cell nucleus (DNA) to cell organelles that are responsible for protein (polypeptide) synthesis. One of the ends in mRNA molecule - called 5’ cap - shows affinity towards the initiation factor present in the cell. Upon their biding, the synthesis of a particular type protein may take place in the cell.

Polish Scientists Solve the Problem For scientists, the main challenge in the development of genetic vaccines was to overcome the natural instability of mRNA molecules which degrade rapidly in cell environment. Without removing this limitation high amounts of mRNA dosed to the cells would be required to obtain a therapeutic effect. Thus, for the cells to produce much higher amounts of desired proteins, mRNA-based therapies would have to ensure both the enhancement of mRNA stability and the affinity to the initiation factor. Only in this way satisfactory therapeutic results might be obtained. And this was achieved by Polish scientists.

mRNA plays an important role in innovative anti-cancer vaccines, cancer therapies, as well as in the treatment of various genetic diseases. mRNA is also one of the foundations in the field of stem cells applications in restoring, forming and treating tissues and organs.

The University of Warsaw cooperated with the Louisiana State University Health Sciences Center at Shreveport (LSUHSC-S). In UW labs the 5’ cap analogues were designed and synthesised, while in LSUHSC-S new mRNA molecules were tested for their stability and affinity in the cell environment.

A Two-in-One Breakthrough

In the cell, natural mRNA molecules are enzymatically degraded (destroyed). Degradation process starts with detaching one of the mRNA molecule ends, so called 5’ cap. The cap is a universal structure which is present in all mRNA molecules of eukaryotic organisms. The scientists from UW focused on the synthesis of 5’ cap analogues which could replace the naturally occurring cap and become more resistant to degrading enzymes activity. Many alternative solutions had been obtained and tested, but only after replacing one of the oxygen atoms with a sulphur atom in a triphosphate bridge, typical for the cap, it was found that the stability of this new mRNA molecule in a living organism extends threefold and, additionally, its affinity towards the factor initiating the protein synthesis increases from 2 to 4 times. As a result, out of the same amount of mRNA over 5 times more protein is yielded. Such a spectacular effect was obtained by exchanging a single atom in a molecule made of 80 thousand atoms. The discovery was conventionally named S-ARCA. Being aware of the impact of their discovery, the same team tested other 5’ end cap analogues and in a short time they found another “effective” analogue, this time by replacing an oxygen atom with a BH3 group. The discovery was conventionally named B-ARCA. Both inventions became a basis to apply for the patent protection.

Solving the mRNA instability issue and enhancing its productivity

The invention has therapeutic applications:

• it is essential in anti-cancer vaccines,

• it facilitates the supplementation of proteins whose low levels cause different diseases,

• it enables reprogramming of stem cells for regenerative medicine purposes.

Before 2007 Studies on the properties of mRNA and 5’ cap analogues

2007 Filing the patent application in Poland and in the USA (S-ARCA); the beginning of work on analogous solutions to protect against competition; publication in a renowned scientific journal

2008 Signing the contract on the co-ownership of the invention between UW and LSUHSC-S (S-ARCA)

2008Filing the patent application in Poland and in the USA (B-ARCA); gaining the interest from research teams seeking potential cooperation; establishing the relation with University in Mainz and BioNTech company - the future investor

2010 Signing the contract on the co-ownership of the invention between UW and LSUHSC-S (B-ARCA)

2010Agreement between UW and LSUHSC-S on the distribution of the revenue from selling the licence; negotiations with the investor (BioNTech); supporting the invention’s credibility by the production and delivery of 5’ cap analogue compound in the amount sufficient to start clinical trials

2010 Signing the licence contract between UW and LSUHSC-S with BioNTech company; first clinical trials set off

2013 Negotiations with BioNTech on the revision of the licence contract due to the global pharmaceutical company interested in the invention which was attracted by BioNTech

2015 Granting by BioNTech the sub-licence to pharmaceutical company Sanofi which conducts clinical trials on a broader scale – a contract worth 360 mln dollars

2016 Selling by BioNTech the sub-licence to Genentech company from Roche group – a contract worth 310 mln dollars

Commercialisation of mRNA 5’ Cap - Timeline

Contract Between the Owners of the Invention After the discovery of 5’-end caps S-ARCA i B-ARCA, UW / LSUHSC-S consortium established and secured with a mutual agreement the co-ownership of the invention. The contract included, among others, the distribution of creative input between institutions, method for managing applications in patent processing, commercialisation process. Due to its considerable experience with the commercialisation process, LSUHSC-S was established a leader for contacting with potential business partners. During next stages both partners also used contracts to regulate issues on the distribution of the revenue from selling licences and sub-licences. The matter is quite relevant as usually in teamwork the business value of patent applications differs, and so does the work contribution or number of co-authors in both applications. Early established terms help to avoid future misunderstandings over the distribution of financial resources between the inventors.

Patent Protection – the Broader, the Better The consortium did realise that in the procedure for patent application it is worth considering the broadest geographical area possible. Applications for patent protection were submitted simultaneously in Poland and in the USA. For both applications the international protection was extended within PCT.

Scientific Publication as an Effective Form of Promotion Immediately after filing patent applications, the team published results of their work in a renowned scientific journal. This was enough to gain the attention from other teams interested in designing and testing “vaccines of the future” which could e.g. permanently treat malignancies. This way UW / LSUHSC-S started the co-operation with a research team from the University in Mainz whose subsidiary, BioNTech company, specialised in research on gene therapies.

Finding a Business Partner BioNTech company was genuinely interested in a licence for new efficient mRNA which would enable conducting the research more effectively, as well as make it possible to introduce life-saving vaccines to clinical trials. BioNTech with their experience in research on genetic vaccines seemed to be a perfect business partner who believed in the potential of the invention and who would invest financial resources in it. For UW and LSUHSC-S it was clear that without such a partner the invention would be of scientific value only. Thus, the negotiations with the future strategic partner began.

Negotiations and Selling the Licence The leader of negotiations (LSUHSC-S) offered BioNTech to prepare a letter of intent outlining the preliminary financial settlements and the strategy for invention protection. However, at the stage of signing the licence contract, a demand was issued to produce 4 grams of the compound containing changed 5’ cap in UW laboratories.

Inventor’s Credibility – Show It Will Work on the Scale BioNTech wanted to minimise the investment risk. By demanding 4 grams of the compound, that is the amount sufficient to start clinical trials, the investor wanted to verify whether the invention, still being in a theoretical sphere, is transferable to the sphere of practical applications. It is worth explaining that such compounds usually are obtained in the amount of 1 - 5 milligrams and the synthesis on the scale expected by BioNTech was about to take weeks. The scientists of UW took on the challenge and after intensive work they proved that they can produce 4 grams of pure 5’ cap S-ARCA.

Gaining the Popularity – Sub-licence and Global Expansion The invention by UW scientists turned out to be groundbreaking enough for the investor to gain the attention of a global pharmaceutical concern and to start negotiations on granting a sub-licence for further clinical trials on a large scale. It required a revision of the original licence contract, including benefits for the inventors and institution. The inventors realised that a bigger player on the market was needed to achieve the implementation of their solution on the global market.

For investors, the full engagement of the inventors in further product development on its every stage is of a great importance. Actually, it was inventors’ engagement and faith in the project that eventually convinced BioNTech to conclude the licence contract.

Commercialisation of mRNA 5’ Cap – Step by step

Head of Gene Expression Laboratory at the Faculty of Physics and Head of Interdisciplinary Laboratory of Molecular Biology and Biophysics at the Centre of New Technologies UW

Since 1980, Prof. Edward Darżynkiewicz has conducted research on the function of 5’ end (so called cap) in eukaryotic mRNA based on synthetic cap analogues; he is a pioneer in this field not only in Poland, but in the world as well (over 170 scientific publications in high level journals, e.g. in Cell. Science, Nature Comm. and the like). In the second half of the 1980s, he initiated synthetic studies on chemically modified dinucleotide cap analogues and on their enzymatic introduction to mRNA in the search for highly translationally active transcripts. His discovery of so called “Anti Reverse Cap Analog”, known as “ARCA”, together with Janusz Stępiński i Robert E. Rhoads (USA) in 2001, occurred to be a groundbreaking invention and has been patented, commercialised and has found a broad application in biotechnological production of proteins. These events have become an inspiration to search for even more effective analogues (beta S-ARCA and beta B-ARCA). In 2014, he was awarded the Leon Marchlewski Medal for outstanding achievements in the field of biochemistry and biophysics.

Edward Darżynkiewicz, Prof.

Head of Laboratory of Bioorganic Chemistry at the Centre of New Technologies UW

He works on the synthesis of nucleotides and their analogues of biological and therapeutic importance. For the last 10 years he has specialised in 5’-end mRNA analogue (cap) synthesis to develop “stable mRNA”, i.e. resistant to cell degradation mechanisms, as well as “effective mRNA” with molecules which stimulate cells to produce certain proteins more efficiently than natural ones. Presently, he is the head of an advanced laboratory at the Centre of New Technologies, University of Warsaw. He has authored and co-authored nearly 100 scientific publications. In his professional career, he has filed 5 patent applications. He is a co-inventor and initiator of two inventions (beta S-ARCA and beta B-ARCA) protected with international patents. Fellow of the Rector Scholarship for years 2004 - 2007, fellow of the Polityka magazine, winner of the UW Rector’s Award for scientific achievements as well as of the Faculty of Physics UW Award.

Jacek Jemielity, Prof. UW

Joanna Kowalska, Ph.D.

Associate Professor at the Division of Biophysics, Institute of Experimental Physics UW

She specialised in the chemical synthesis and studying properties of modified nucleotides of biological importance. For the last few years she has worked on the design and synthesis of 5’-end (cap) mRNA analogues to apply in biotechnology and medicine. She has authored and co-authored more than 50 scientific works. Fellow of, among others, the Minister of Science and Higher Education Scholarship, Foundation for Polish Science. Many times awarded for scientific achievements, e.g. with the UW Rector’s Second Degree Award, the Faculty of Physics UW Award, Prof. Pieńskowski Award. She is a co-inventor of two inventions (beta S-ARCA and beta B-ARCA) protected with international patents.

Co-inventors of beta S-ARCA and beta B-ARCA from UW

Co-authors of the invention from UW

Co-authors of the invention from LSUHSC-S

Joanna Żuberek, Ph.D., Assistant Professor at the Division of Biophysics, Institute of Experimental Physics UW, co-inventor of beta B-ARCA.

Maciej Łukaszewicz, Ph.D., Assistant Professor at the Division of Biophysics, Institute of Experimental Physics UW, co-inventor of beta B-ARCA.

Ewa M. Grudzień-Nogalska, Doctor of Philosophy (PhD), Biochemistry, Biophysics and Molecular Biology.

Robert E. Rhoads, Professor, Head of the Department of Biochemistry & Molecular Biology.

What you should know about UOTT University Technology Transfer Centre is a part of University of Warsaw and it helps scientists to commercialise their work (discoveries and inventions). UOTT supports scientists throughout the whole commercialisation path – from the idea, through filing a patent application, to establishing a spin-off company, selling a licence or a single disposal of rights.

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