Cynthia A. Challener is a contributing editor to Pharmaceutical Technology.

Cynthia A. Challener

Numerous trends in the pharmaceutical industry, and particularly in the biopharmaceutical sector, are leading to the need for smaller-capacity production facilities, which in turn is affecting the types of investments being made by pharmaceutical manufacturers. First is the shift to biologic-based drugs, which has occurred along with a dramatic increase in the productivity, mainly in terms of much higher titers, of biopharmaceutical processes. “While demand has increased for biopharmaceuticals, it has not kept pace with the 10-fold increase in productivity,” states Günter Jagschies, senior director of strategic customer relations for BioProcess, GE Life Sciences. “As a consequence, there is an opportunity to scale down biologics manufacturing, and newer facilities are typically based on 2000-L or smaller reactors and are much less cash-intensive, which is a key factor for pharmaceutical companies today given the growing pressure for them to be more cash aware,” he adds.

The second factor is the shift to production in emerging markets, both because the middle class is increasing and demand for drugs is expected to grow significantly, and in response to individual government requirements, such as in Brazil and Russia, for western manufacturers that want to sell products--both large and small-molecule--in those countries to produce the drugs locally. “At present, however, demand for biopharmaceuticals in particular remains limited, generally due to affordability issues, so smaller production volumes are still required. In addition, domestic manufacturers are already serving these markets, which further reduces the required facility size,” Jagschies notes. In the biosimilar sector, the stiff competition will also keep volumes quite low, at least initially. There are approximately 100 companies worldwide looking to produce 5-7 of the most successful biologics that are coming off patent, and even if just 10 of them are successful, each will have a small share of the market, according to Jagschies. At the same time, many new drug candidates (i.e., antibody drug conjugates) have much higher potency and thus require much smaller doses, which also translates to lower production quantities. 

Increasing access to single-use technologies for biologics production is further enabling the scale-down of manufacturing facilities, because such systems provide flexibility and enable companies to quickly change their portfolios in response to market needs. For batch reactions in particular, Jagschies notes that the use of disposable technology can also help improve cash flow. The introduction of modular facilties is another enabling factor. “With modular facilities, companies can export the technology required for quality production to anywhere in the world, including areas where it would not be possible to construct such facilities otherwise,” says Jagschies. By their nature, modular facilities are smaller-scale.

“All of these trends together are driving interest in smaller facilities and smaller investments, particularly in the biopharmaceutical sector,” Jagschies concludes.

A summary of selected investments in both small- and large-molecule manufacturing facilities during the last year is presented as follows.

: Abbott signed an agreement in June 2014 to acquire Russian pharmaceutical manufacturer Veropharm for approximately $495 million, according to the company. Abbott will acquire Garden Hills, the holding company that currently owns more than 80% of Veropharm, but is expected to own more than 95% by the time the transaction closes. Veropharm’s portfolio includes products in the areas of women’s health, central nervous system disorders, cardiovascular disease, gastroenterology, and oncology. Veropharm is currently constructing a new manufacturing facility to expand its production capabilities, according to Abbott.

: AbbVie officially opened its newly expanded facility in Sligo, Ireland in June 2014 (announced in 2012), which provides manufacturing capacity to deliver on AbbVie’s existing portfolio of medicines and to support potential new therapies within its pipeline, according to the company, which invested €85 million ($115 million) in the project. AbbVie also announced in February 2014 that it will invest $320 million to build a facility in Singapore for the production of both small-molecule and biologic APIs. The facility will have 250 workers once it is operational in 2019, according to the company.

: Actavis is investing $48 million to reopen in Puerto Rico its solid dosage manufacturing and packing facility in Manati and expand hormone production at its site in Fajardo. The company expects to create 300 jobs by the time the facilities are operational in 2016.

: AstraZeneca announced in late 2013 that it is expanding its presence in Macclesfield, UK. The company is investing $190 million in a new plant to increase production of its blockbuster prostate cancer drug Zoladex (goserelin acetate). While no new jobs will be created, the decision does mean that the existing 300 jobs at the site will be retained.

: Bayer HealthCare is making several investments at home in Germany and abroad. The company will spend nearly $694 million at existing sites in Leverkusen and Wuppertal, Germany to expand production of its hemophilia drug Kogenate (antihemophilic factor), adding 500 workers by 2020. Bayer Healthcare is also investing $138 million to expand its packaging, logistics, and analytical capabilities at its facility in Beijing, China.

: After investing more than $350 million to improve manufacturing operations for injectable drugs at its Ben Venue Laboratories site in Bedford, OH, Boehringer Ingelheim announced that the plant would be closed because the cost and time required to overcome systemic problems was too high. Across the globe in China, the $95 million expansion of its plant in Shanghai that began two years ago and included the addition of manufacturing, packing, and distribution operations is now operational. Boehringer Ingelheim also indicated that it would make further investments at the site in 2014 to establish a development lab for biologic drugs.

: BMS is consolidating more than 575 jobs in information technology, marketing services, and financial services at its new North America Capability Center over the next three years. In 2013, the company also announced a $250 million expansion of its biologics plant in Devens, MA, adding R&D and clinical trial manufacturing capabilities. Over in Ireland, however, it is closing an API plant near Dublin and eliminating one manufacturing line at its API plant in Swords County. 

: The $450 million, 240,000-square-foot biologics facility in Kinsale, Ireland that Eli Lilly announced in 2012 is expected to be complete by the end of 2014, with the first production of GMP lots taking place in early 2016. In 2013, the company also announced nearly $1 billion in planned plant expansions for the production of its insulin products, including API and cartridge manufacturing capabilities: $245 million at its sites in Indianapolis, IN and Puerto Rico, $120 million in France, and $350 million in China. In addition, Lilly is spending $60 to $70 million on the construction of a 260,000-square-foot manufacturing facility in Nantong-Jiangsu, China with its Chinese partner Novast that will produce generic versions of several Lilly drugs. This plant is expected to be operational by the end of 2014.

: GSK and Novartis made big news in April 2014 when they announced a three-way swap of various businesses. GSK is selling its oncology business to Novartis and buying Novartis’ vaccines business (excluding influenza vaccines), and the two companies are combining their over-the-counter businesses.

In March 2014, GSK announced plans to invest as much as £130 million ($222 million) in five African manufacturing facilities and an R&D laboratory for non-communicable diseases over the next five years. Possible locations include Rwanda, Ghana, and Ethiopia. In the same month, the company announced that it is increasing production of several over-the-counter products at is Zebulon, NC plant. In September, 2013, GSK opened a $40 million plant in Cork, Ireland for the production of Gantrez, the adhesive used in the company’s Poligrip denture product.

Also in 2013, following its announcement in 2012 that it would invest more than $790 million upgrading two production sites in Scotland and construct a new plant in Ulverston, Cumbria, GSK indicated that it would invest a further $330 million on additional upgrades and a new technology center focused on continuous manufacturing, biocatalysis, nanotechnology, and supply-chain processes. The company also intends to use continuous processing for some of the manufacturing processes at the new facilities. 

In November 2013, GSK announced that it will invest approximately $136 million to build a new facility in India that will also use continuous processing. The facility is expected to be operational in 2017. The company is also constructing a $50 million facility in Singapore that will use continuous processing.

: Jannsen, a business unit of Johnson & Johnson, announced plans to construct a $200 to $300 million, 267,000-square-meter plant in Xi’an, Shaanxi province, China to replace a Janssen Pharmaceutical facility built by J&J and a Chinese partner in 1985. The new facility will serve as a supply-chain center for China and other Asian countries.
Merck KGaA: The company’s biopharmaceutical division Merck Serono will construct an €80 million ($108 million), 431,000-square-foot facility for the bulk production and packaging of drugs for the treatment of diabetes, cardiovascular disease, and thyroid disorders in Shanghai, China that is expected to be completed in 2016 and ready for production in 2017. In Europe, Merck Serono is spending $68 to $69 million (€50 million) to modernize its fill-finish plant in Bari, Italy. Merck’s Allergopharma unit, meanwhile, is constructing a $55 million (€40 million), 64,600-square-foot facility near Hamburg, Germany for the production of allergy medicines destined for China and other emerging markets. It is expected to be completed in 2016. 

: In addition to its purchase of GSK’s oncology business and sale to GSK of its vaccines operations (except influenza vaccines) in April 2014, Novartis also announced that the two companies would combine their over-the-counter businesses in a joint consumer-health venture.  At the same time, Novartis announced that it was selling its animal-health business to Eli Lilly. According to the company, it now has a more focused portfolio with depth in dermatology, heart failure, respiratory, cell therapy, and oncology in its pharmaceuticals business, a strong position in eye care through its Alcon unit, leadership in generics with its Sandoz business, and is well-positioned for innovation with Novartis Institutes for BioMedical Research (NIBR).

Also in April 2014, Alcon announced the opening of a S$200 million (US$160 million), 330,000-square-foot plant in Singapore for the production of products for the treatment of dry eye, allergies, bacterial infections, and glaucoma. In addition to the $500 million the company announced in 2012 that it would spend on a second facility for biologic drugs, Novartis indicated in February 2014 that it is expanding one of its existing plants in Singapore, adding a new GMP warehouse and raw material laboratory. 

Novartis is also working with African partners to expand research capacity in the region.  In 2013, Novartis conducted workshops in Kenya and Ghana to help African researchers construct safe and effective Phase I trial facilities for bioequivalence, and may hold additional workshops in other countries. NIBR and the University of Cape Town in South Africa also launched a collaboration in 2013 to expand research capabilities at the university’s Drug Discovery and Development Center.

In December 2013, Novartis announced the closure of an outdated Ciba Vision plant in Canada.

Novo Nordisk: Novo Nordisk is constructing a $100 million, 29,100-square-foot purification pilot plant at its new research and development facility in Denmark to expand its capacity for early-phase diabetes projects. The facility is being built to allow for a doubling of capacity if needed. The company is also constructing a plant in Denmark with R&D centers for diabetes and biologics, and recently announced plans to add 6000 positions in Denmark by 2022, half in R&D and half in production.

 Pfizer recently completed a $30 million expansion of its product technology lab in Ireland and is spending an additional $100 million to upgrade its biologics plant in the country. 

: In Ireland, Regeneron is spending $300 million to create a pharmaceutical plant at a former 400,000-square-foot Dell computer manufacturing site and expects to employ a total of 300 people by 2016. The company is also investing $70 million to complete the second phase of an expansion of its East Greenbush, NY facility, which will include the addition of two 10,000-liter bioreactors.

: Following its 2013 announcement to expand its biologics production capabilities at a cost of more than $880 million, Roche said in April 2014 that it will also be upgrading its small-molecule manufacturing operations, investing approximately $135 million to construct a new plant at is headquarters in Basel and improve other facilities for the production of both developmental and existing drug products. The new plant is expected to be onstream late in 2016.

The biologics investments include ~$210 million for a new ADC manufacturing plant in Basel, ~$286 million to expand production capacity at two sites in California, and nearly $385 million to upgrade its Penzberg, Germany plant.

Roche is also spending $105 million on a new packaging and logistics training center in Switzerland.

 Sanofi is investing in emerging markets. The company is not only increasing insulin production at its facility in Russia, it is investing $95 million in a new manufacturing plant in Algeria, which will have a capacity of 100 million units per year, and $75 million to construct its third facility in Vietnam, which will be operational in 2015. Meanwhile, Sanofi’s Genzyme business is investing $80 million at its recently approved facility in Framingham, MA, adding more downstream processing capabilities for Fabrazyme (agalsidase beta), its treatment for Fabry disease.

: Teva’s $2 billion cost-cutting efforts continue and will include the definite closure of 11 plants, and possibly an additional 16, which are under evaluation by the company. Plans to build an API plant in India have also been scrapped. In October 2013, however, Teva opened a $100 million plant in Croatia that produces tablets, capsules, and sterile formulations for the European and US markets. 


 Cynthia Challener is a contributing editor for 

Articles

Shrinking facility size, growth of biologics, and emerging market demand influence pharma investments.

Pharma Investments Reflect Key Industry Trends

Continuous processing for the production of key building blocks and intermediates for small-molecule APIs is no longer viewed as a technology of the future. Most pharmaceutical companies with in-house manufacturing use flow chemistry to some extent, and smaller companies that outsource production expect their contract-manufacturing partners to have continuous-flow systems available. Advances in microreactor technology for commercial-scale production and implementation of continuous downstream processes will ultimately enable the complete continuous synthesis of complex organic molecules required for small-molecule API manufacturing. Widespread adoption, however, will occur slowly as the industry shifts from the infrastructure in place today to smaller, modular, and flexible facilities.


“Continuous flow synthesis is a great alternative to traditional batch synthesis/production when it comes to demanding chemistries such as hazardous reactions or potentially challenging conditions like high pressure and temperature,” observes Dominique Roberge, head of continuous flow/microreactor technology business development at Lonza Custom Manufacturing. In addition, continuous flow processing provides many technical advantages over traditional batch methods, including speed of development, process reliability and product quality, maximization of process safety when employing hazardous chemistries, and minimization of investment during product development, according to Peter Poechlauer, principle scientist at DSM Fine Chemicals. For Jörg Schrickel, new business development manager for CABB, the more consistent quality that is achieved with continuous manufacturing, particularly for sensitive products, is of significance.

 “There are, in fact, some products that cannot be produced in large batch processes due to the thermodynamic nature of the reactions involved. Under continuous flow conditions, however, because only small quantities of reagents and products are present at any given time, these issues can be avoided,” Schrickel adds.

Another advantage is the improved environmental footprint of continuous processes. Poechlauer points to reduced raw material consumption and waste generation as two key benefits. CABB increases the sustainability of its continuous processes for the production of acid chlorides and derivatives by recycling off-gases (referred to by the company as its “verbund and recycling system”). “As a result, our building blocks have very low process mass intensity (PMI) and environmental factors (E-factors),” says Schrickel.


Despite the obvious technical advantages of continuous flow processing, companies are adopting the technology steadily, but slowly, and in a strictly opportunistic way, applying elements of continuous processing technologies when they pay off immediately,” states Poechlauer (1). “The technology is not developed to a certain degree of maturity and then waits to be applied, but is developed for and in close connection with a certain production process,” he adds.

Flow chemistry is often considered as an option when exploring new chemical routes, increasing yield, lowering the cost of goods, and generating IP, according to Roberge. “One of the main drivers for the implementation of flow chemistry is, however, to perform a reaction that cannot be done with traditional batch production, such as those involving azide intermediates or oxidation using oxygen,” he notes. Currently, the drivers for adoption are academia and institutes and providers of continuous flow equipment who are looking for innovation and often work together with the pharmaceutical industry to solve specific problems, according to Schrickel.

The main issue is the large existing batch manufacturing infrastructure. “If a company gets trapped in the question ‘use existing or build new?’, it will use the existing equipment in the present business environment,” states Poechlauer. Continuous equipment like microreactors are added when needed, and the existing vessels modified to serve other purposes, such as hold-up tanks used to define the batch for regulatory purposes. “We expect most pharmaceutical syntheses will continue to be a mix of continuous and batch manufacturing operations, which also enables manufacturers to build in inventory buffer zones,” observes Poechlauer.

In one recent example, researchers at GlaxoSmithKline found that for the large-scale manufacture of potassium bromomethyltrifluoroborate, a key raw material for a Suzuki−Miyaura coupling reaction (2), a hybrid approach involving the use of both continuous and batch processing where most appropriate was successful in achieving the project goals, according to Toby Broom, a chemist with GlaxoSmithKline R&D.


“The pharmaceutical industry is very much interested in continuous processes, and a lot of activity is underway within most pharmaceutical companies related to the development of continuous processes. However, we do see continuous processes being more relevant for the next generation of APIs,” says Schrickel. Lonza believes that at least the top 20 pharmaceutical companies have flow systems within their assets at this point in time, with some of the larger players calling continuous processing a disruptive or breakthrough technology, according to Roberge. At the same time, small to mid-size pharma and biotech companies often evaluate flow technology through outsourcing. “As a result, contract manufacturers that have flow processes in place are better positioned to engage in partnerships and collaboration discussions with these early start-ups,” Roberge comments. 

As the internal advocate groups within pharmaceutical companies push for the development of continuous processes, continuous processing will be applied in a growing number of processes, but in an evolutionary way, rather than a revolutionary one, as people come to accept the technology as mainstream rather than “experts only” territory, according to Poechlauer. In addition, he notes that with large pharmaceutical houses focusing on new leads rather than on process development, custom manufacturing organizations continue to play an important role in this field. 

Support from the authorities for continuous processing has also been crucial in furthering its acceptance by small-molecule API manufacturers. “The industry and FDA are in constant dialogue on continuous processing,” Poechlauer says.

Advances in continuous commercial production


One of the biggest issues for continuous flow chemistry has been scaling up to commercial production levels, because most early equipment for flow chemistry was designed for the laboratory. According to Poechlauer, however, chemical manufacturers and pharmaceutical companies have been working with microreactor manufacturers to address this issue, and pilot- and small commercial-scale equipment is now available, often by “numbering up” and running several microreactors in parallel. “One advantage of this approach is that modular designs that can be fitted to the specific needs of different reactions are possible,” he says. 

This advantage was taken into consideration when DSM installed its commercial-scale microreactor suite. “We are very interested in microreactor technology and continuous processing because, in some cases, it makes it easy to scale up reactions that can’t easily be done otherwise due to the nature of the raw materials or the kinetics of the reaction,” says Poechlauer. 

Lonza has designed a new type of production facility concept called the “Factory of Tomorrow” in which FlowPlate microreactors of various sizes are incorporated into the company’s development and manufacturing plants. “Flow processes lead to high intensification that radically reduces the size of a reactor and its footprint,” Roberge states. The range of FlowPlate equipment allows for production of a few grams of product in the lab setting and up to 5-ton campaigns for later-stage projects. “In such an environment, flexibility and versatility is needed to combine novel process conditions at high temperature and pressure, which can be difficult to achieve in a classical plant setting. Continuous manufacturing can significantly improve process efficiency with a reduced footprint, therefore reducing overall material consumption,” he observes.

In addition to addressing scale-up issues, advances have also been achieved in expanding flow chemistry beyond liquid systems, which are the traditional and most simple systems for continuous processing, according to Schrickel. “Much progress has been made with respect to the development of technology for the continuous processing of liquid/solid and multi-phase systems,” he explains. Lonza, for example, is developing microreactor plates with its partner Ehrfeld Mikrotechnik BTS, a Bayer Technology Services company, that will enable multi-phase reactions. Schrickel believes these advances will be important for the further application of continuous processes in API production. 

Poechlauer does add that there are uncertainties about scale-up, and managing intellectual property issues must be done carefully, given that in many cases new process chemistry is involved. 


More recently the focus has been on integrating continuous downstream processes with continuous flow synthesis. “Most downstream processes can be easily adapted to run continuously; extractions, phase separations, and distillations are actually more effective when done continuously,” says Poechlauer. On the other hand, he notes that solids handling processes, such as filtration and crystallization, present some difficulties. There are solutions being introduced to the market, however, such as belt filters and spray-drying, and Poechlauer expects others will follow.

Lonza, in fact, is currently extending the application of continuous flow technology to address continuous work-up unit operations such as distillation, according to Roberge. “The objective is the continuous operation of a complete mini-plant within the manufacturing unit ‘Factory of Tomorrow,’ which will allow its use for larger-scale manufacture,” explains Roberge.  


Research and development of continuous processes will continue and grow, first on specific, isolated topics, because there might not always be a solution that will enable a complete continuous process or at least a process without bottlenecks. To switch to continuous processes, companies will have to gain all of the possible benefits that the technology offers, and sustainability will be one of the key aspects, asserts Schrickel.

Poechlauer also believes that continuous processes will ultimately lead to simplified distributed manufacturing (on-site, on-demand) in modular container plants as well as enable application of “personalized medicines.” He also notes that the use of disposable devices in the manufacture of highly potent compounds will be beneficial to the further development of continuous processes for these challenging compounds.

“Our partners in the pharmaceutical industry are in a transition stage and are moving toward integrated solutions. Laboratory chemists consider the environmental consequences of raw materials and processes when developing synthetic routes. Drug manufacturers are also forming closer collaborations with suppliers to explore new technologies and concepts, such as continuous processing and the use of microreactors. Such relationships will help demonstrate the benefits of this approach and ultimately lead to adoption of process intensification when it is the most effective solution,” Poechlauer concludes.

At the same time, continuous processes will require new equipment and perhaps dedicated production lines for each API, according to Schrickel. “Pharmaceutical companies may insource the entire manufacturing of their APIs or force their outsourcing partners to install dedicated continuous plants. In either case, the adoption of continuous processes may result in a radical change in the pharmaceutical and outsourcing industry,” he asserts.

 17 (12), 1472-1478 (2013). 
2.	T. Broom et al., Org. Process Res. Dev. Article ASAP, DOI: 10.1021/op400090a Publication Date (Web): June 10, 2013.


Cynthia A. Challener is a contributing editor to 

The pharma industry is moving toward commercial-scale continuous processes for small-molecule API manufacturing.

The Mainstreaming of Continuous Flow API Synthesis

Unlike in the chemical and food industries, and even in small-molecule pharmaceutical manufacturing, which have well-established methods for automated online sampling, the biopharmaceutical sector remains in need of a reliable system that transfers bioprocess samples directly from bioreactors to analytical devices while maintaining process sterility. Such a system is necessary if biopharmaceutical companies are to effectively take advantage of process analytical technology (PAT) and gain a more fundamental understanding of what is happening within bioreactors.  While on-line Raman and dielectric spectroscopy provide some insight, there remains a need for integration of this data with off-line measurements, such as cell density, viability, metabolite levels, and titers. 

To overcome this issue, Bend Research (part of Capsugel Dosage Form Solutions), in collaboration with major biopharmaceutical companies, including Pfizer, Eli Lilly, and Boehringer Ingelheim, has developed the Modular Automated Sampling Technology (MAST) platform. MAST consists of a sterile sampling system, a sample scheduler and navigator for automated sampling and delivery of samples to various analytical instruments, and a data-management system for collecting and analyzing the results, and if desired, providing feedback to the bioreactor for direct adjustment of reaction conditions.  


The heart of the MAST platform is the new sterile sampling unit referred to as the Sample Pilot, which consists of a sampling module and an associated programable logic controller (PLC). There are two Sample Pilot modules, the SP100, which collects 50-mL samples for monitoring of pilot- and commercial-scale manufacturing operations, and the SP200, which takes incremental 5-mL samples for analysis of development-scale reactions and reactions that run in smaller laboratory-scale bioreactors and pilot-scale, single-use bioreactors. Each sample assembly consists of proprietary valves and a pump and is designed to draw samples from the bioreactor and push them out to user-defined destinations, according to Clint Pepper, a director at Bend Research. “Because we use positive displacement, rather than pulling the samples out with a vacuum, it is possible to deliver even viscous culture samples with high cell densities to analytical instruments more than 50 feet away.”  

Importantly, both systems can be sterilized in place (SIP), autoclave-sterilized, or gamma-sterilized depending on the model selected. “The entire system can be cleaned and sanitized between each sampling, and there is flexibility for users to adapt various cleaning procedures depending on their specific needs,” notes Lisa Graham, senior vice-president of Bend Research.   


To maximize the potential of sterile biopharmaceutical sampling, the MAST platform design includes software systems for managing sample scheduling, delivery of the samples, processing of the analytical results, and adjustment of bioreactor parameters if desired. The MAST Sample Scheduler and Navigator modules will work together to facilitate sample collection from bioreactors and transfer to analytical testing systems. 

The MAST Data Management system will maintain information on sampling activities and analysis results and has the ability to automatically adjust the feed rate of the bioreactor to meet desired growth rates according to the current condition of the cells. “With this data management system, it is possible to significantly reduce the time needed to process large sets of analytical results. Information on the state of cells and the process is obtained more rapidly, ” Pepper asserts. 

On-line measurement combined with at-line analysis  


Early studies at Pfizer have shown that automated at-line measurements obtained using the MAST system are similar to those obtained when using manual samples. The MAST platform has been incorporated with on-line analytical technologies to deliver overall cell-level “observability” and “guidance,” according to Graham. In one case, dielectric spectroscopy measurements were corrected to more accurately estimate total and viable cell density. Off-line measurements of Caspase-3 activation were related to on-line measurements of the variation in the capacitance due to the reduction in cell viability following staurosporine addition. This information was used to determine cell health in the bioreactor in real time. In another example, the amount of off-line data that was collected (Raman spectroscopy) was greatly increased due to the ability to more frequently collect samples automatically and without the need for an operator. The data were used to build robust process-monitoring models and develop a Raman system for laboratory use, according to Graham. “In this case,” she observes, “the Sample Pilot serves as enabling technology for the implementation of PAT.” 


The steam-sterilized SP100 module has been successfully demonstrated in five different bioreactors in three different facilities. “During recent development runs on pilot-scale bioreactors, the MAST platform and SP100 successfully delivered more than 480 samples without contamination, including more than 200 cumulative days of successful operation at the 30-L to 500-L scales in both cell culture and microbial applications,” reports Pepper. “The sanitant-sterilized SP200 module has been deployed in five different facilities ranging in scale from bench reactors to large-scale, single-use bioreactors. The SP200 units have collected over 2000 samples in more than 240 accumulative days of operation with no contamination or loss of sample integrity.”

MAST systems have been placed in nine client facilities (15 total units installed) with more than 2400 sterile samples collected without loss of sterility.  “The major MAST product modules have moved out of the beta testing stage and are being sold to early adopters as pre-commercial products,” says Pepper. 


New MAST technologies are still being beta tested, including a module to automatically remove cell from the whole broth, leaving the clear supernatant. This module would send clear samples to analytical devices that require cell-free samples, such as liquid chromatographs and mass spectrometers, so that these instruments can be integrated for real-time data collection and processing. 

In the near future, Bend Research plans to install the first multiplexed MAST system capable of collecting samples from up to eight bioreactors and sending those samples to as many as four analytical devices. “This solution should gain a lot of traction in the laboratory setting where banks of bioreactors are running at the same time,” says Pepper.


The MAST platform offers the ability to collect reproducible samples more frequently and automatically without operator involvement, which reduces the risk of contamination and operator exposure. The ability to integrate analytical results from numerous sources provides more insight into cell behavior and the impact of process parameters, enabling the use of more sophisticated process-control strategies.  

“Our intention with the MAST platform is to help biopharmaceutical companies gain insight into what actually occurs in a bioreactor by providing a means for understanding how bioprocess variables affect overall product quality,” comments Graham. “We believe that by coupling the right tools, the MAST platform provides in-depth guidance for optimization of the reactor environment in order to meet cell needs for the production of the target product. Ultimately, we hope that the MAST platform will provide the meaningful real-time analytics, process measurements, and novel data management techniques that will enable the development of predictive control models that in turn will drive increased product consistency and batch-to-batch reproducibility,” she concludes. 

Cynthia A. Challener, PhD is a contributing editor to 

The Modular Automated Sampling Technology platform allows sampling of bioreactors.

Automating Online Sampling of Bioprocesses

Reliable systems to transfer bioprocess samples directly from bioreactors to analytical devices while maintaining process sterility are crucial for biopharmaceutical companies to effectively take advantage of process analytical technology (PAT) and gain a more fundamental understanding of what is happening within bioreactors.  
   
  Bend Research (part of Capsugel Dosage Form Solutions) has collaborated with major biopharmaceutical companies, including Pfizer, Eli Lilly, and Boehringer Ingelheim, to develop the Modular Automated Sampling Technology (MAST) platform. MAST consists of a sterile sampling system, a sample scheduler and navigator for automated sampling and delivery of samples to various analytical instruments, and a data-management system for collecting and analyzing the results, and if desired, providing feedback to the bioreactor for direct adjustment of reaction conditions.

The MAST platform includes the Sample Pilot, a sterile sampling unit that consists of a sampling module and an associated programmable logic controller (PLC). The SP100 Sample Pilot modules collects 50-mL samples for monitoring of pilot- and commercial-scale manufacturing operations, and the SP200 takes incremental 5-mL samples for analysis of development-scale reactions and reactions that run in smaller laboratory-scale bioreactors and pilot-scale, single-use bioreactors. Each sample assembly consists of proprietary valves and a pump and is designed to draw samples from the bioreactor and push them out to user-defined destinations, according to Clint Pepper, a director at Bend Research. "Because we use positive displacement, rather than pulling the samples out with a vacuum, it is possible to deliver even viscous culture samples with high cell densities to analytical instruments more than 50 feet away."

To maximize the potential of sterile biopharmaceutical sampling, the MAST platform design includes software systems for managing sample scheduling, delivery of the samples, processing of the analytical results, and adjustment of bioreactor parameters if desired. The MAST Sample Scheduler and Navigator modules will work together to facilitate sample collection from bioreactors and transfer to analytical testing systems. 


  The steam-sterilized SP100 module has been successfully demonstrated in five different bioreactors in three different facilities. "During recent development runs on pilot-scale bioreactors, the MAST platform and SP100 successfully delivered more than 480 samples without contamination, including more than 200 cumulative days of successful operation at the 30-L to 500-L scales in both cell culture and microbial applications," reports Pepper. "The sanitant-sterilized SP200 module has been deployed in five different facilities ranging in scale from bench reactors to large-scale, single-use bioreactors. The SP200 units have collected over 2000 samples in more than 240 accumulative days of operation with no contamination or loss of sample integrity."   MAST systems have been placed in nine client facilities (15 total units installed) with more than 2400 sterile samples collected without loss of sterility.  "The major MAST product modules have moved out of the beta testing stage and are being sold to early adopters as precommercial products," says Pepper. 

The MAST platform offers biopharmaceutical manufacturers several advantages, according to Pepper, including the ability to collect reproducible samples more frequently and automatically without operator involvement, which reduces the risk of contamination and operator exposure. The ability to integrate analytical results from numerous sources provides more insight into cell behavior and the impact of process parameters, enabling the use of more sophisticated process-control strategies.  

Cynthia Challener, PhD, is a contributing editor to 

This article was excerpted from an article that will appear in the July issue of 

The Modular Automated Sampling Technology platform takes online samples from bioreactors and transfers them directly to analytical devices.

Advances in Real-Time Bioreactor Sampling

Advances in catalysis have had a tremendous impact on the pharmaceutical industry, enabling the synthesis of highly complex small-molecule intermediates and APIs in fewer steps, with increased atom economy, and often with high selectivities and yields. There are issues with many of the most widely used catalytic systems, however, including the cost, availability, and toxicity of many of the precious metals used to form the catalysts and the need for expensive and complex ligands to achieve the desired transformations. There has been much interest, therefore, in the development of alternative catalysts derived from abundant, nontoxic metals and simple ligand systems. New base metal catalyst platforms are also of interest because they may enable new chemistry not previously observed with traditional precious metal catalysts. Select examples with potential applications in pharmaceutical synthesis are discussed below.


  There is a finite supply of metals in the Earth’s crust, and humans are consuming many of them at an ever-increasing rate. As a result, the availability of certain transition metals for use in the production of catalysts is becoming increasingly limited due to growing demand for these materials in other applications, including the newer organic light emitting diode displays in smart phones and televisions and renewable energy systems, such as fuel cells, wind energy, and photovoltaics. 

In fact, in 2012, the British Geological Survey updated its risk list for the supply of chemical elements or element groups that are of importance to human society (1). The availability of each element was determined by considering its abundance in the Earth’s crust, the location of current production and reserves, the political stability of those locations, and the recycling rate and substitutability of the element. The rankings give an indication of which elements might be subject to supply disruption, most likely from human factors such as geopolitics or resource nationalism, along with events such as strikes and accidents. At the top of the list with the highest risk of supply disruption are the rare earths, platinum group metals, niobium, and tungsten. It should be noted that the demand for the elements (outside of their substitutability) was not considered in this analysis.

The platinum group metals are of particular interest to the pharmaceutical industry, because many important catalysts rely on platinum, palladium, ruthenium, rhodium, iridium, and osmium. These metals are expensive and experience high price volatility. The price of platinum, for example, ranged from $1400-$1500/ounce in March 2014, and has varied more than 5-fold in the last few years, according to Paul Chirik, the Edward S. Sanford Professor of Chemistry at Princeton University. With respect to platinum supply, he notes that the amount of platinum produced each year is only approximately 6% of the quantity of gold mined annually.


  Some researchers are raising the question of whether or not a reaction can truly be considered sustainable if such rare (and at times toxic) metals are required. According to Amir Hoveyda, Joseph T. and Patricia Vanderslice Millennium Professor of Chemistry at Boston College, even if a transformation is otherwise green (i.e. minimal/no solvent, high atom economy, no need for heating/cooling, little downstream processing requirements), it can’t be realistically considered as sustainable if a catalyst based on a rare transition metal is required. Hoveyda also notes that to be industrially practical, reactions must proceed in a reasonable period of time, ideally within six to eight hours, and catalysts must provide high selectivities and high yields.


  One reaction that Hoveyda has focused on is the copper-catalyzed synthesis of enantiomerically enriched homoallylic amines and alcohols, which are important intermediates for the preparation of many different biologically active molecules. As an alternative, his group has developed a transition metal-free route to enantiomerically pure amines and alcohols with inexpensive, easily prepared derivatives of the amino acid valine as catalysts for the reaction of a wide range of readily available unsaturated organoboron reagents with imines and carbonyls. In addition to not requiring any rare transition metal salts, the reactions are generally complete in six hours, require low catalyst loadings, and provide high yields and enantioselectivities. The effectiveness of this new set of catalytic reactions, according to Hoveyda, is due to the use of a proton and internal hydrogen bonding.


  Chirik, meanwhile, is developing alternative catalysts based on cobalt and iron, which are far more abundant than the precious metals. In addition, while most precious metal catalysts require two-point substrate binding for inducing high levels of enantioselectivity in asymmetric hydrogenation, because iron and cobalt are a new catalyst type, they have yet to exhibit this limitation, he points out. For example, Chirik’s cobalt asymmetric hydrogenation catalysts formed using simple and inexpensive chiral amine ligands are relevant to the pharmaceutical industry. In fact, Chirik developed the catalysts through a collaboration with Merck &  Co. that included high-throughput experiments. Notably, the cobalt catalysts are effective for the hydrogenation of both functionalized and unfunctionalized olefins in high yield and high enantiomeric excess. 

In a second example, Chirik’s group has developed iron catalysts that mediate both inter- and intramolecular [2π + 2π] reactions of alkenes and alkynes to form cyclobutenes. What is interesting about this system is that it is effective for unactivated olefins and is also selective for 2+2 cycloaddition even in the presence of a diene that can undergo Diels-Alder (4+2) cycloadditions. According to Chirik, such selectivity is unique to iron and results from its ability to engage in radical chemistry with the supporting ligand.


  A third strategy for the development of alternative catalysts involves mimicking the behavior of enzymes. Such research is being performed by Robert Morris, a professor at the University of Toronto, who has developed new iron catalysts that perform what he calls “metal-ligand bifunctional catalysis,” in which the ligand and metal both play an active role in the catalytic cycle. Notably, he has shown that with the right choice of ligands, the electronegativity of the iron center can be adjusted such that it is close to that of carbon and thus able to form covalent bonds, particularly iron-hydrogen bonds. Surprisingly, carbon monoxide, which is often a poison to catalysts, is a required ligand as it is for nature’s hydrogenases.

Such catalysts are effective for the asymmetric transfer hydrogenation (ATH) of ketones and can be simply prepared from relatively inexpensive diamines and phosphine aldehydes, which self-assemble (promoted by the metal through a template effect) to form imines that bind to the metal. Morris notes that the catalyst is effective at low loadings and the iron can be readily oxidized upon exposure to air and then precipitated, allowing for simple purification. A third-generation family of these iron-based catalysts was reported in 2013 and should be available for purchase soon (2).  These catalysts are more active than any known catalyst for ATH and as selective as more expensive conventional ruthenium-based systems.

The catalyst may also be applicable for direct hydrogenation under a mild hydrogen pressure, and the reverse of the asymmetric transfer hydrogenation reaction may be useful for the kinetic resolution of racemic alcohols.


  Carbon-carbon coupling reactions have become widely used in organic synthesis, including the commercial manufacture of numerous pharmaceutical building blocks and key intermediates. Robin Bedford, professor of catalysis at the University of Bristol, has been investigating the potential for using iron catalysts to mediate these important transformations. In the past, coupling reactions involving iron have generally been Grignard reactions of “hard” nucleophiles. Bedford wanted to expand the repertoire to other “softer” nucleophiles (e.g., zinc) that are used in coupling reactions catalyzed by platinum and palladium.

What his group found was that the choice of ligand was crucial for creating iron catalysts that are effective for both the Negishi coupling of benzyl halides and phosphates with diarylzinc reagents and Suzuki couplings of arylboronates with alkyl and allyl halides and 2-halopyridines. Most importantly, the best ligand is the simple, inexpensive, bis(diphenylphosphino)ethane (dppe) ligand. The key, according to Bedford, is to prepare the catalyst in the correct manner. “One of the important lessons we learned in this work was that it is important to understand the identity and role of the active catalytic species and how ligands interact with iron in order to generate the desired species,” Bedford states.


  Iron is not generally thought of as a good metal for the preparation of catalysts because of its electronic structure. Platinum group metals, which are heavier metals with no unpaired electrons, have always been thought to form stronger bonds with ligands, according to Patrick L. Holland, a professor of chemistry at the University of Rochester. On the contrary, his group has shown that complexes based on metals with unpaired electrons can follow similar reaction mechanisms, and often do so more rapidly, as long as a balance is maintained between the weakness and activity of the ligand-metal bonds. 

Holland has developed special bidentate ligands that are designed to afford the high-spin electronic configuration in complexes with first-row metals. For example, his group has developed an iron(III) imido complex that oxidizes hydrocarbons and transfers a nitrene (NR) group to organic substrates.

“We believe that properly designed complexes of iron and other first-row metals can serve not only as alternatives to transition metal catalysts for the mediation of established reactions; given their unusual reactivity, they may also provide access to novel transformations,” observes Holland.

“Risk list 2012: An updated supply risk index for chemical elements or element groups which are of economic value,”

 Accessed Apr. 10, 2014.
  2.	W. Zuo et al., Science, 342 (6162), 1080-1083 (2013). 

Industry seeks alternatives to costly and toxic precious metals used to form the catalysts for organic synthesis.

Alternatives to Precious Metal Catalysts Sought

Transition metal-containing catalysts are widely used in organic synthesis, but many are based on expensive, rare metals. Researchers are seeking more sustainable alternatives.