Asian innovation: a unique opportunity for the pharmaceutical industry. Fall 12
As a rapidly developing economy, China is an important emerging market that offers incredible new opportunities to pharmaceutical companies seeking to advance R&D and drug development productivity. A unique opportunity exists to build on China’s strong history of innovation by investing in research and development facilities that will keep the pharmaceutical industry at the forefront of scientific advances, today and in the future.
One particular company, AstraZeneca, recognised the potential of the Asian market several years ago, and has made significant investment in the region to position itself to benefit from this growth. The company’s Wuxi site, its largest manufacturing site in Asia, was established in 2001 at an initial cost of $134 million, followed by a further $35 million investment five years later to increase capacity at the site. Since then, there has also been considerable additional investment in formulation – $50 million – as well as the establishment of a Regional Packing Centre and High-Tech Receiving Centre. In research and development, AstraZeneca is forging ahead with innovative drug discovery studies relating to the Asian population. In addition to this significant financial investment, the importance of developing local research collaborations was recognised, and extensive efforts have been made to build strong relationships with biotech companies, contract research organisations (CROs), hospitals and academia, plus individual researchers and entrepreneurs. Today, the company employs more than 4,700 people in China who are actively involved in manufacturing, sales, clinical research and new product development.
At the forefront of drug discovery
In recent years, Asia has become one of the fastest growing markets for the pharma industry, yet innovative pharmaceutical research and development in China has been quite limited. However, this gives companies the advantage of starting with a blank canvas, leaving them free to pioneer new ways of conducting research, rather than duplicating Western R&D models.
In June 2006, AstraZeneca’s senior executive teams announced a $100 million strategic investment in China to establish a translational research centre, the AstraZeneca Innovation Center China. The ICC was initially set up as a function group focusing on translational science for oncology research, with an emphasis on diseases that are prevalent in the Asian population. Liver, gastric and lung cancers are particularly common in the region; the World Health Organisation estimates that 78% of liver cancer patients and 74% of gastric cancer patients worldwide are in Asia, and that mortality rates are also higher in this area. As a global translational science centre, the ICC’s remit during the first three to four years was to explore the company’s portfolios – in particular, the oncology portfolio – and investigate disease indications in Asian patients that could be due to ethnic differences. With the pharmaceutical market in China and Asia continuing to grow, further investment followed and, over the next few years, the ICC evolved into a drug discovery centre covering early drug discovery screening, pre clinical and early clinical development. Today, its mission is to deliver candidate drugs and proof of concepts, helping to produce new medicines to address the previously unmet medical needs of Asian patients.
By establishing the ICC in Shanghai, AstraZeneca has positioned itself to take advantage of China’s vast pool of scientific talent. Currently, around 80 scientists are employed at the ICC and with staff from a wide variety of scientific backgrounds – from academia to the pharma and biotech industries – the ICC benefits from an extensive range of experience in a wide range of different disciplines. While many of the ICC’s scientists have been trained in China, others have gained additional experience by working overseas, acquiring a different perspective on drug discovery. This introduces diversity and external know-how to the work of the ICC.
In addition to harnessing the talents of its own scientific staff, the ICC actively seeks innovative R&D partners throughout China and Asia. The formation of long-lasting relationships with key opinion leaders, hospitals, clinicians, CROs, industry and academia has enabled the ICC to build an established research network and embark on collaborations that benefit translational science and drug discovery worldwide. These collaborations allow the ICC to complement its own core expertise with external knowledge and capabilities, enhancing its efforts to make groundbreaking discoveries and develop new medicines faster.
Moving drug discovery forward
As a pioneering drug discovery centre, the ICC features a number of key departments including chemistry, bioscience, translational science and medical science, with each department subdivided according to its expertise and function. Like many other large pharmaceutical companies, the ICC relies on a matrix model, where departments combine their capabilities and expertise for effective project management. A team comprising scientists from various departments is formed at the commencement of a study and a project leader – or sometimes two project leaders from different disciplines – is appointed to manage the team. This allows the project team to draw on a range of expertise for all the design, planning, experimental work, data interpretation and decision-making relating to the study.
Each department is responsible for the delivery of its daily workload and will select a protocol to meet its individual needs. The chemistry department relies on its core capability to design a study and implement quality control, and also frequently collaborates with contract research organisations, which are well placed to deliver the experimental data. Bioscience and translational science provide data through a number of different inhouse laboratories, including in vitro and in vivo pharmacology, drug metabolism pharmacokinetics (DMPK) and histopathology, using state-ofthe- art equipment and technology platforms. High throughput screening can also be accommodated if the need arises.
The ICC’s drug discovery process centres on developing a detailed knowledge of the genetic mechanisms of disease, and the determination of clinically-relevant biomarkers and drug targets that are related to the Chinese and Asian populations. Once a disease area has been identified, the ICC works closely with hospitals and doctors, with the ultimate aim of developing therapies that are targeted to Asian patients. The search for suitable drug targets begins with a detailed literature review that ensures the disease segment is fully understood. Collaborations with hospitals follow, which are crucial to surveying patient samples for the targets. In this respect, the ICC’s Shanghai location plays a key role, allowing vital relationships to be fostered with key opinion leaders in the field, as well as clinical centres, in the battle to understand a disease and to identify targets. Working in partnership with hospitals allows extensive disease profiling to be performed and, subsequently, a list of suitable drug targets to be identified. Target validation is performed in-house and once the list has been reduced to just a couple of priority targets, the drug discovery screening process is launched. This approach has already seen success in the development of new therapies for gastric and liver cancer, with new compounds progressing into Phase I and II clinical trials, and the ICC plans to extend its research into other areas, such as metabolic and respiratory diseases.
Supporting drug discovery with innovative screening techniques
A critical part of the drug discovery process is candidate compound screening to identify novel drug candidates, and scientists at the ICC continually seek to develop original techniques – and to improve existing methods – to streamline this process. One such example is the development of an automated 3D cell-based assay. 3D cell-based assays have been used in academic cancer research for the past decade, and have the potential to provide a better model to simulate the growth and formation of tissues in the body. However, the technique does have limitations for drug discovery screening due to low throughput and reproducibility issues. Scientists at the ICC successfully adapted a 3D cell culture assay to enable it to be carried out on an industrial scale, after automating assay plate preparation on a liquid handling platform. This allowed 3D cell-based assays to be performed in 96-well microplate format.
Matrigel™ (BD Biosciences) is a gelatinous protein mixture secreted by Engelbreth-Holm-Swarm (EHS) mouse sarcoma cells, which induces the growth of cell colonies in a different manner to the standard cell culture surface (Figure 1). It contains structural proteins that provide the adhesive peptide sequences that cultured cells would encounter in their natural environment, as well as growth factors that promote cell differentiation and proliferation and, when used as a thick three-dimensional gel, induces cell migration from the surface to the interior of the gel.
Matrigel’s heterogeneous composition confers the ability to stimulate complex cell behaviour that would normally be difficult to observe under laboratory conditions. The production of colonies that mimic the growth and formation of tissues in the body, as well as the migratory behaviour, make it ideally suited to the study of tumour cell-cell contact, growth, migration and metastasis, aiding researchers in the identification of the right compound during early phase drug discovery.
Increased throughput and consistency with automation
Historically, the ICC has performed Matrigelbased 3D cell growth assays in six-well plates. Manual plate generation for the assay is both problematic and time-consuming because of the large volumes needed and the complex handling requirements of the Matrigel. As large volumes of reagents and candidate compounds are required, scaling up the assay is difficult to achieve, making this format poorly suited to drug screening; there was a clear need to optimise the assay for costeffectiveness. A further problem is that Matrigel is extremely difficult to pipette in low volumes, as its heterogeneous composition leads to high surface tensions and uneven distribution across a well, making the analysis of results extremely difficult, and virtually impossible for 96-well plate format. The ICC resolved the issue by developing a protocol to pre-coat the microplate wells with a thin layer of soft agar prior to adding the Matrigel.
The challenge remained to introduce automation to enable medium throughput compound screening and improve consistency. Although the surface tension issues associated with the media had been resolved, the use of an agar layer complicated pipetting logistics; agar solidifies below 50°C, while Matrigel becomes too viscous to pipette above 4°C. The Innovations Center decided to take advantage of the flexibility and modularity of Tecan’s Freedom EVO® 150 liquid handling platform, which was already used for compound serial dilutions and a number of other cell-based assays. Temperature-controlled carriers were added to the workstation to allow both the agar and Matrigel reservoirs to be kept at appropriate temperatures for pipetting. Because of the workstation’s modularity, it could be adapted quickly and economically without interfering with other applications, making it the obvious choice for automation of the Matrigel assay.
All pipetting is performed using a single channel on the system’s Liquid Handling (LiHa) Arm; the risk of blockages is avoided by pre-heating or precooling the tip by multiple aspiration and dispense cycles. Hot agar, at around 55°C, is pipetted into pre-warmed 96-well plates using the LiHa Arm’s multi dispense function. This minimises the risk of premature cooling of the media and allows small media reservoirs to be used, as well as helping to further reduce costs. After allowing the plate to cool at room temperature on the deck of the instrument for 15 to 20 minutes, a Matrigel layer can be added to the now solid agar. The automated protocol enables the preparation of six complete plates in one and a half hours, compared with only three or four plates in an entire working day manually. The quality of the plates has also improved tremendously, eliminating the batch-tobatch, plate-to-plate and even well-to-well variation that often occurs with manual plate preparation, providing the consistency that is vital for drug screening applications.
But preparation of the Matrigel plates is just one stage of a procedure that also involves cell seeding, compound stamping and clone checking (Figure 2). The Matrigel plates are transferred to a Matrix® WellMate® and Microplate Stacker (Thermo Scientific) for cell seeding and, after incubation, returned to the Freedom EVO for compound stamping using the MultiChannel Arm™ 96 (Tecan), taking advantage of the flexibility offered by advanced automated platforms. Further incubation is followed by the addition of reagents prior to performing readings with either a Safire2™ (Tecan) or an EnVision® Multilabel Reader (Perkin Elmer). In addition, cell counting is performed using a Guava® cell counter (Merck Millipore), and an IX71 inverted microscope (Olympus) is used for cell observations such as clone checking. This streamlined process ensures that the ICC’s scientists obtain the high quality results required to help accelerate the drug discovery process, highlighting the importance of continued investment in state-ofthe- art laboratory solutions to keep the ICC at the forefront of research and development.
For the ICC, location is the key and, geographically, it is important for the Center to be based in China. China is now the third-largest market for the pharma industry, and increasing numbers of pharmaceutical companies are opening centres in the China/Asia region. With its remit to focus on diseases with a high prevalence in Asia, it was vital for the ICC to be situated close to relevant patient populations, physicians and hospitals, enabling collaborative studies to be undertaken to understand each disease. The ICC’s Shanghai location has allowed it to develop mutually beneficial relationships with other interested parties and provided access to key opinion leaders. This supports exploration of existing AstraZeneca compounds in the disease areas or patient segments of interest, as well as the identification of new targets, enabling innovative drug discovery programmes to be launched. With new and enhanced methods developed by talented scientists, the Innovation Center China is perfectly positioned to fulfil its mission, launching increasing numbers of targeted therapeutics specifically designed for the Chinese and Asian populations.
Dr Yi Gu is a Director of the AstraZeneca Innovation Center China (ICC) and leads the translational science group. Her group provides preclinical data and biomarker analysis to support personalised healthcare strategies for candidate drugs. Dr Gu is one of the founding members of the ICC, helping to established its scientific capabilities in 2006. She had 15 years of experience in translational research in the US before joining AstraZeneca in China.
William Shi, MS, is a Senior Automation Specialist at the ICC. He provides automation, instrumentation and assay support for compound management, HTS screening, cell panel screening and projects in drug discovery and translational sciences. Before joining the ICC in March 2008, he worked in academia and industry in China for seven years.