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BIA-Brukerstyrt innovasjonsarena

Development of a new production process for a last resort antibiotic

Alternative title: Utvikling av en ny produksjonsprosess for et siste skanse antibiotikum

Awarded: NOK 16.0 mill.

Project Manager:

Project Number:

296372

Project Period:

2019 - 2022

Funding received from:

Subject Fields:

The project is a collaboration between Xellia Pharmaceuticals AS, and SINTEF with Leiden University as a subcontractor. The aim is to develop a new manufacturing process for an antibiotic with several advantages compared to related products such as one-time dosing, higher antimicrobial activity, and activity against antibiotic resistant bacteria (MRSA). The antibiotic is made by a bacterium isolated from soil that produces the compound in very low amounts. To make an economical competitive process it is crucial to develop mutants of the bacterial strain with significantly increased ability to produce the active ingredient. Furthermore, a fermentation process, a purification process as well as a process for chemical synthesis, that converts the natural antibiotic into the finished product, must be developed. There is during the project established high throughput methods for isolation of mutant strains with improved antibiotic production, including effective protocols for introducing random mutation into the cell’s DNA, miniaturized fermentation conditions and rapid analyses making it possible to analyze a high number in a short time. This has made it possible to isolate new mutants with significantly increased levels of antibiotic production that can be applied in an economical industrial process. The improved mutants have been genome sequenced and compared to the wild-type strain to identify mutations that contribute to the increased production. Roughly 50-100 mutations were found in each strain per generation. In some of the improved strains mutations were observed in the regulator genes for the antibiotic coding genes, while in other overproducing strains it has been challenging to identify the mutation(s) that most likely are contributing to the improved production abilities. At Leiden University it has been establish molecular methods for site specific mutagenesis in the wild-type strain using CRISPR-cas9 technology. The idea is to combine several beneficial mutations and avoid additional mutations with negative impact usually seen for classical mutants. By use of these methods some of the mutations identified to be the cause of increased production have been introduced to the wild-type strain resulting in improved production levels. In the last part of the project a considerable amount of time was spent on improving the fermentation process to reduce the level of some troublesome impurities not removed by purification. A new fermentation process based on the new production strains as well as new processes for chemical synthesis and purification has been scaled up to pilot and full production scale. Production of product within specifications has been demonstrated. Further work will include to run three subsequent validation batches, develop the finish formulated product and commercialize the product.

- New production process for Dalbavancin, developed with basis in the N. gerenzanensis wild type strain to secure freedom to operate for Xellia Pharmaceuticals AS. - Improved generic product portfolio for Xellia after product launch to market in 2025, important both for the company and for a secured supply of important MDR antibiotics to large patient groups globally. - Proof of concept for a novel strategy for strain development based on genome sequencing, bioinformatics, and genetic engineering using developed CRISPR-Cas9 technology for Actinobacteria. - A list of mutations correlated to improved productivity in the wild-type strain. - Increased competence in pharmaceutical industrial strain and process development in Norway.

The project is a collaboration between Xellia Pharmaceuticals and SINTEF with Leiden University as sub-contractor. The aim is to develop a new manufacturing process for a last resort antibiotic with several advantages compared to related products such as beneficial pharmacokinetics, higher antimicrobial activity and activity against antibiotic resistant bacteria (MRSA). Xellia has the ambition to be the first company to enter the generic market when patents have expired. This requires development our own proprietary production process; i.e. the production strain, fermentation and purification processes, chemical modification as well as finished dosage formulations compliant with regulatory authorities such as FDA and EMA. Launch of the generic antibiotic will make the product available for larger patient groups world-wide at significantly reduced costs. A new approach for rapid strain and process development will be pursued. Xellias API R&D capabilities have been located in Oslo for more than 50 years, and the site employs 32 scientists/engineers, specialized in classical strain development, fermentation, synthesis and purification technologies. Xellia is a trusted provider of life-saving anti-infective treatments and are supplying antibiotics and finished dose formulations to more than 500 pharmaceutical companies in 70 countries. Xellia has state of the art production facilities with global regulatory approval. SINTEF Industry, department of Biotechnology and Nanomedicine located in Trondheim has the knowledge, experience and infrastructure for DNA sequencing, advanced mass spectrometry analysis, process and cultivation studies, as well as high throughput screening. The SINTEF group is currently leading/partner in several international projects focusing on biopharmaceuticals and drug development. Leiden University, The group of Professor Gilles van Wezel, are world leading experts on genetic engineering of Streptomyces and rare actinobacteria.

Funding scheme:

BIA-Brukerstyrt innovasjonsarena