Working together to expedite biologics development and improve availability: the production of pre-clinical grade biopharmaceuticals

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By Bernard Massie and Denis Bourbeau

The pharmaceutical research paradigm is rapidly shifting from the traditional approach of screening for small chemical entities as potential drugs toward the development of biologics targeting novel untapped molecular targets and new mechanisms of action. In fact, it is estimated that 300 biologic products are in clinical trials, with another 600 to 700 in pre-clinical or early clinical development. When it comes to drug targets, there are an estimated 1,800 possible targets for protein-based therapeutics. Furthermore there are an estimated 2,100 possible targets for gene therapy1. With their relative insulation from generic competition combined with their ability to address unmet medical needs and novel targets biologics undeniably represent the most attractive therapeutic modality for the new era of molecular medicine.

While Canada ranks second after the United States for the number of biotech firms2, it relies twice as much on universities to perform research as the U.S. does3. This means that the bulk of the research in Canada places more emphasis on the R than on the D of R&D. In the Industry Canada document ‘The Canadian Biopharmaceutical Industry Technology Roadmap,’ it is reported that companies are started prematurely, before they have a chance to sufficiently develop high quality IP. This is in part because Canadian universities have limited technology transfer opportunities and thus rely on new company formation as a technology transfer strategy, as opposed to the U.S. where there are both financial support and a critical mass of pharmaceuticals companies to act as receptors4.

The report concludes that a major weakness in Canada is the relative lack of activity in new biological entities, and the lack of access to the needed technologies and proper investments. The lack of risk capital for drug development is a significant barrier; currently VC money is often used for IP protection rather than product development. This is where the NRC is stepping in; it is generally recognized that the needed structure for applied research is quite distinct from university’s discovery research labs. For a successful technology transfer, one needs to be timely and efficient; this means protecting and strengthening IP, testing proof-of-principle, and assessing the market. It is NRC’s mandate to do just that. With its organizational structure, networking and wide range of research activities, NRC scientists have the means to regroup into dedicated multidisciplinary teams to move technology forward.

NRC mandate and involvement
in biologics

At the NRC, we are undertaking, assisting, and promoting scientific and industrial research in different fields of importance to Canada such as the Pharma/Biotech sector. In this latter sector, NRC is providing national access to integrated R&D and to innovation support through mainly two institutes (BRI and IBS). With its enabling technologies and unique facilities, it plays a significant role in the Canadian biotechnology sector by facilitating the translation to the clinical arena of findings from academic and industrial research laboratories as well as its own.

In the past few years, a multidisciplinary team of NRC scientists has concentrated their effort on the development of anti-cancer biologics and have produced several promising molecules. Just recently, it has entered into a licensing agreement with Alethia Biotherapeutics for its monoclonal antibody (mAb) against Clusterin, an antibody primarily targeting tumour invasion. Earlier, the same team had transferred a single domain antibody to Helix Biopharma, another Canadian company. This single domain antibody was combined with the company’s proprietary technology and is now preparing to enter human clinical studies. When companies license out NRC’s technologies, they are not left alone. Typically both organizations keep working hand in hand to further develop the technologies. Both Helix and Alethia, for instance, have entered into several collaborative agreements to characterize the therapeutic moieties in animal models and to humanize the monoclonal antibodies.

Recently, the NRC has made substantial investments in order to increase the throughput of its mAb facility. It is now capable of increased mAb production through the combination of multiplexed immunization and roboticized high-throughput hybridoma screening. It can also conduct relatively high-throughput label-free binding studies as well as functional studies for rapid validation of the mAbs. This is one example of NRC’s commitment in positioning itself to help support new technology discovery and maturation.

Biotherapeutics: From
discovery to developability

The NRC is further reducing the gap in technology maturation by providing infrastructures and expertise for the optimization and validation of the biologics it develops and those of its partners. Namely, it has leading edge technologies in bioprocessing. NRC scientists and their partners have access to an integrated biotherapeutics production facility: from cDNA to 1500 L. Its teams of scientists have developed over the years proprietary expression systems in prokaryotes (M. extorquens, E. coli), and in mammalian cells (CHO, 293, NS0). To facilitate the transition toward clinical trials, it has large-scale production and purification methodologies under GLP, for ensuring a smooth transition to cGMP.

From cDNA to protein in
three weeks

Producing biologics in mammalian systems currently requires stable transfections, typically in CHO cells, and the selection of high producer clones. This process requires 16 to 24 weeks and there are several inconveniences with this approach: 1) the generation of stable clones is a lengthy and cumbersome process, 2) one needs to maintain a cell line for every candidate protein under study, 3) revival of a frozen high-producing clone sometimes proves to be difficult with expression being lost, and 3) there is no flexibility; if one needs to introduce a change in the protein sequence one has to go back to square one.

In contrast, protein production by transient transfection can be done in less than three weeks, and only one cell line needs to be maintained. However, yields are typically low, transfection reagents are expensive, and the need to change media is cumbersome and costly. NRC scientists have developed a method of transient transfection that does not require an expensive transfection reagent nor media change, that is fully scaleable, and that can generate protein at high yields. This technology has attracted a large number of industrial partners and clients.

Viral vectors for gene therapy and vaccination form another very interesting class of biotherapeutic; this technology is the one that has the potential to tackle the largest number of therapeutic targets. When it comes to gene therapy, NRC scientists have been onboard for more than a decade. Its teams of scientists have not only developed novel platforms that carry great promise in the field of cancer and vaccines, but have also invested in industrial scale vector production and purification.

Earlier this year, NRC’s scientists and their collaborators reported in Cancer Research the success of their viral platform in significantly improving the efficacy of an adenoviral vector carrying a suicide gene for cancer therapy. This vector had an improved activity through increased suicide gene activity and heightened immune response in immunocompetent mice. This platform also holds great potential for new vaccines. NRC’s multidisciplinary team has also developed leading edge technologies for the large-scale production (up to 500 L bioreactor) and purification of viral vectors in Biosafety Level-2 facilities.

NRC’s unique organizational model has empowered its scientists to build multi-disciplinary teams in protein- and viral-based biotherapeutics development. While developing its own IP, NRC is also co-developing enabling technologies that address some of the bottlenecks in the process. With unique expertise and facilities to support the ability of Canadian SMEs to move forward the commercialization of biotherapeutics, the NRC hopes to bridge the gap that hinders Canadian innovation from making its way efficiently to market.

References

1.     The Canadian Biopharmaceutical Industry Technology Roadmap, Industry Canada
2.     Global Biotechnology report 2006, Ernst & Young
3.     The Canadian Biopharmaceutical Industry Technology Roadmap, Industry Canada
4.     The Canadian Biopharmaceutical Industry Technology Roadmap, Industry Canada

Bernard Massie is the director, Bioprocess sector at the Biotechnology Research Institute, National Research Council in Montréal, QC.