Jennifer Markarian is manufacturing reporter for Pharmaceutical Technology.

Jennifer Markarian

The Brazilian rainforest is one of the world’s most biodiverse regions and a center for phytochemistry, in which indigenous plants are harvested and their chemicals extracted for use as pharmaceutical APIs. Optimizing the process of gathering, harvesting, and extracting phytochemicals depends on the use of sustainable harvesting practices and modern business and manufacturing processes.

	Centroflora CMS, a joint venture between phytochemicals expert Centroflora from Brazil and the Swiss contract manufacturing organization (CMO) CMS Pharma, specializes in phytochemical processing. In 2015, the company was recognized for the way it handled its acquisition and technical transfer of Boehringer Ingelheim’s (BI) non-captive phytochemical API portfolio in 2014.  This work won the company CPhI’s 2015 Award for Excellence in Partnering and Outsourcing, which was presented at CPhI Worldwide in Madrid in October 2015.

	Andrew Badrot, founder and CEO of Centroflora CMS, discussed the project’s scope and challenges with 

 What were some of the reasons BI chose Centroflora CMS?

 Securing this complex business and executing the technology transfer was crucial because several of the APIs involved are used to make life-saving medicines. The chosen partner would be responsible for ensuring supply continuity but would also have to deliver additional services and support for related products. Centroflora CMS offered three benefits: phyto API manufacturing capability, the outsourcing expertise required to manage the API technical transfer, and the ability to take over all business activities.

 What were some of the keys to ensuring the successful technical transfer of BI’s API phyto product portfolio?

The main challenge for Centroflora CMS was to establish an acceptable fast-track process that would allow it to transfer products, customers, and all the services associated with an API production site change, and to do all these things concurrently. Centroflora CMS set up the proper infrastructure and transferred manufacturing know-how from BI in record time. We have also taken over more than 130 customer relationships worldwide. This relatively smooth transition provided the basis for continuity of market supply of all the essential-medicine APIs involved in the transaction.

	Centroflora CMS’ project management team worked to transfer the products in a methodical way, based on documentation provided by BI. By soliciting limited input from BI’s technical team and by relying mainly on existing documentation, we shielded experts at BI from the substantial resource commitments that are typical for technical transfers.

	On the receiving end, a CMO with a wide range of chemical engineering and regulatory experience worked out innovative manufacturing solutions and allowed the project to be completed in only 10 months from start to finish.

 Can you discuss how you worked to secure the supply of the API, pilocarpine?

 CMS, through its partner Centroflora Brazil, secured the vital market supply access route for pilocarpine, an important API used in the treatment of glaucoma and dry mouth disease. This success was based on the cooperation of many stakeholders within the framework of Centroflora’s program called 'Partnerships for a Better World.' Sponsors of this program also included BI and the German government institution 'Gesellschaft für Internationale Zusammenarbeit' (GIZ).

	The partnership helps indigenous people in Brazil harvest Jaborandi (the leaf containing pilocarpine API) in a sustainable way. The project uses fair-trade principles and ensures a stable source of income for approximately 1000 indigenous families who live in the Brazilian rainforest. The partnership has demonstrated that big and small companies can cooperate successfully to achieve a common goal-supplying patients with APIs in the most effective way while ensuring that all stakeholders remain committed and engaged.

 What are your plans for expanding this program?

 First, we are interested in sharing the 'Partnerships for a Better World' program with other companies that harvest natural products. Second, integrating other phyto-derived APIs into the program is a strategic objective to help grow a biodiverse API portfolio, which, in turn, should pave the way to find and develop new, sustainable pharmaceutical products.

	Other organizations have recognized the potential shown by this business model. In 2015, the Union for Ethical Biotrade, a non-profit organization dedicated to promoting sustainable development and biodiversity, granted approval to Centroflora. Group members commit to sustainable sourcing practices and are required to undergo periodic audits.

	The Brazilian government recognized us for both environmental and social efforts. The National Confederation of Industry in Brazil named Centroflora’s pilocarpine harvesting program one of the country’s 10 most sustainable business practices. The Bank of Brazil also recognized the firm for its work to improve conditions for farmers in the northern forest region of the country.

	We look at the pilocarpine success story as the way forward for combining bio-based API business with 'fair trade' concepts around harvesting. Ideally, other pharmaceutical companies will be inspired to follow suit. 'Partnerships for a Better World' is an ecological and socially responsible approach to harvesting botanicals. It secures the ability to bring patients around the world a portfolio of trusted, well-established, and vital medicines in a sustainable and responsible way. The program, together with our large investments in API manufacturing with an FDA-approved factory in Brazil, provides the elements needed to attract new phyto API businesses to Brazil. 


	"Partnership Strategies in Outsourcing"
	

	When referring to this article, please cite it as J. Markarian, "Success Story: Transferring a Phytochemical API Portfolio," in 

“Partnership Strategies in Outsourcing” 

Articles

Sustainable harvesting combined with CMO expertise helped Centroflora CMS ensure supply continuity after it acquired Boehringer Ingelheim's non-captive API phytochemicals portfolio.

Success Story: Transferring a Phytochemical API Portfolio

	Pharmaceutical companies are increasingly making public commitments to sustainability goals and investing in "green" chemistry and the equipment and manufacturing practices needed to meet these goals. For example, in November 2015, Patheon, Biogen, Johnson & Johnson, Genentech, and Novartis, signed the American Business Act on Climate Pledge and declared their goals, which include reducing carbon or greenhouse gas emissions, water use, and waste to landfill, and increasing the use of renewable energy (1).

	Using cleaner energy generation is one way to address these targets. Power plant company FuelCell Energy, for example, announced plans to install a combined heat-and-power fuel-cell power generation system at Pfizer’s Groton, CT R&D facility to provide low carbon electricity and steam. The installation is expected to be operational by summer 2016 (2).  

	Key components of a “green manufacturing” program are reducing waste, energy, and water use, said David Lynch, who heads up environment, health, and safety, and engineering for GlaxoSmithKline (GSK)’s global manufacturing and supply in an 

. The company has reduced energy consumption by investing in cleaner energy generation, renewable energy, and using waste for energy, for example. The company cut water use in its operations and is working with suppliers to reduce their water impact.  GSK is working toward its goal of halving hazardous and non-hazardous operational waste by 2020 through programs to first eliminate waste, and then reuse, recycle, or generate energy from waste (3).

	AstraZeneca China employed sustainable manufacturing design and practices at its solid-dose facility in Taizhou, China, which won the International Society for Pharmaceutical Engineering (ISPE) 2015 overall 

Facility of the Year Award for its Taizhou Supply Site Project

. Optimizing the process and minimizing waste are primary sustainability issues that can be addressed through programs such as lean design, minimizing energy usage, optimizing plant footprint, and preventing failures in the manufacturing process that result in un-usable product, commented Martin Teo, Project Director of the Taizhou Supply Site Project at AstraZeneca, in an interview with 

. In the Taizhou project, an area of focus was optimizing the utilities that surround the manufacturing process, such as water (purified water and water for injection) and heating, ventilation, and air conditioning (HVAC). For example, in the first year of data collection, over 10% of the annual utility costs were saved using an HVAC setback system that automatically reduced air changes during non-operational hours. “We have designed a manufacturing facility which truly supports the manufacturing process, and transport and material wastes are minimized. Best practice tools such as material flow reviews, materials staging, and spaghetti diagrams were used to make the ‘wastes’ clearly identifiable,” explained Teo.

	Preventing release of API into the environment was another area of focus for the Taizhou project. For Betaloc (metoprolol) production, for example, the team identified an innovative electro-oxidation process to pretreat Betaloc-containing waste water prior to conventional biological treatment. Oxidation converts the toxic API into smaller, non-toxic molecules and removes more than 99% of API from the waste water, reports AstraZeneca. The process is fully automated and the cost and environmental impact of running the system are minimal. The company notes that the process copes well with fluctuations in API concentration.

	With innovations such as these, scientists and engineers are on their way to making a greener future for pharmaceutical manufacturing.

	References
	1. “Pharma Companies Join the American Business Act on Climate Pledge,” Dec. 3, 2015, 

www.pharmtech.com/pharma-companies-join-american-business-act-climate-pledge

	2. “FuelCell Energy to Install Clean Power at Pfizer R&D Facility,” Jan. 13, 2016, 

www.pharmtech.com/fuelcell-energy-install-clean-power-pfizer-rd-facility

Pharma companies reduce the impact of manufacturing on the environment with innovations in energy and waste treatment.

Pharmaceutical Manufacturers Go Green

The pharmaceutical industry has an important role to play in implementing solutions to global environmental challenges, such as the availability of clean water and reducing energy use. GlaxoSmithKline (GSK) is one example of a healthcare company that is making significant strides toward reducing its carbon footprint through “green” manufacturing. A company’s carbon footprint is a measure of its environmental impact that is determined by calculating the greenhouse gases emitted during sourcing, production, use, and disposal of the company’s products, stated in terms of equivalent tonnes of carbon dioxide (CO

). In-house operations, including manufacturing and laboratories in GSK’s pharmaceutical, vaccine, and consumer health divisions, accounted for approximately 11% of the company’s carbon footprint in 2014, says the company. GSK is well on its way to meeting ambitious goals it set to reduce energy and water use, as well as waste, in its operations and across its value chain, in addition to its long-term goal of being carbon-neutral (i.e., removing as much CO

from the environment as it emits) by 2050. David Lynch, who heads up environment, health and safety and engineering for GSK’s global manufacturing and supply at GSK, spoke with 

 about some of the projects GSK has undertaken in the manufacturing area to achieve its sustainability goals. 

 What do you see as the key components of a ‘green manufacturing’ program, particularly looking at your direct operations and your own facilities? 

 Reducing energy, water, and waste in direct operations are important elements of a green manufacturing program. GSK Manufacturing (GMS) has reduced energy consumption by 47% and water consumption by over 55% since 2001. Progress has been achieved by a number of significant initiatives focused on our processes, utility infrastructure, and buildings, where a significant proportion of consumption resides.  We use lean sigma, operational excellence, and standard work techniques to ensure that a consistent, efficient procedure is identified and locked in for routine operation. By standardizing at the level of best performance, savings are achieved and propagated by sharing them as best practice.

	We are investing in energy efficiency, cleaner energy generation (combined heat and power), renewable energy (solar and wind), using waste for energy, and reducing the amount of coal we consume by 60%. We have installed wind turbines at our Barnard Castle (UK), Irvine (Scotland), and Cork (Ireland) sites. We have installed 13 combined heat and power, natural-gas engines in the UK and Europe. We are in the process of designing and building a biomass steam-raising plant in Ireland.

	The lessons learned from improving the efficiency of our existing factories are now being embedded in any new facilities; we conduct rigorous, multi-discipline, sustainability reviews of new designs before the investment is approved. The scope of such reviews includes water, waste, sustainable materials of construction, energy efficient lighting, air conditioning motors, and heat and cooling systems, for example.

 What have you identified as the most significant areas of waste in pharmaceutical manufacturing, and how are you reducing waste?

 We categorize waste into hazardous and non-hazardous waste. In pharmaceutical manufacturing (as opposed to the manufacturing of consumer healthcare products), management of hazardous waste is the biggest challenge. For small molecules, the principal waste is hazardous organic solvents. That said, because solvents are so valuable, we try to recycle and reuse as much of the solvent as we can. Where we can’t either recycle or reuse, we have implemented projects to incinerate waste solvent to generate energy at our factories, most notably in Singapore and at Cork in Ireland.

	Larger molecules don’t tend to use large amounts of hazardous solvents, so we take other approaches to recycling any waste generated. For example, we have received regulatory approval to compost 1500 tonnes per year of the egg waste we generate from manufacturing flu vaccines at our Sainte Foy site in Canada. Instead of sending it to landfill, the egg waste will be mixed with other green waste and sold as compost.

	In GSK overall (i.e., our pharmaceutical, vaccine and consumer health operations),  our goal is to halve operational (i.e., non-hazardous and hazardous) waste by 2020. We are doing this by adopting four simple steps to eliminate, reuse, recycle, and generate energy from waste-in that order of priority. We are making steady progress. In 2014, we generated 159,000 tonnes of waste from our operations, which was 4% less than in 2013 and 11% less than 2010, and half of our manufacturing sites and major R&D sites are now sending no waste to landfill.

	The proportion of waste that is recycled or disposed of with a positive benefit has also increased from 71% in 2010 to 74% in 2014. We continually look for innovative ways to reduce and recycle waste at other sites around the world in conjunction with our waste management partners. For example, we send packaging waste from our Poznan site in Poland to make a construction material for waterproof flooring.

 What are some of the best practices you use to reduce hazardous waste?

 Since 2010, six of our pharmaceutical manufacturing sites have recycled and re-used 126,000 tonnes of organic solvents. Specifically, at our Jurong, Singapore site, we have installed boilers that will burn up to 10,000 tonnes per year of waste solvent emanating from both the Jurong site and the nearby Quality Road site. This process generates steam that is used to generate chilled water for use in the air conditioning systems at the Jurong site. This initiative has enabled Jurong to achieve a 50% reduction in hazardous waste. In addition to the environmental benefits, this project has also helped us build trust and has further enhanced our reputation in Singapore. At the 2014, 17th Singapore Environment Achievement Award (SEAA), organized by the Singapore Environment Council, the Jurong site won two prestigious manufacturing awards.

	Perhaps the most impact has been achieved by our Quality Road site. At one time, 24% of its waste was hazardous, but this level has now been reduced to < 4% with an in-house treatment using photo-oxidation technology to reduce the high levels of chemical oxygen demand (COD) and phenol.  This technology reduces COD by 40% and thus contributes towards a greener manufacturing process.

What are some ways you have reduced water usage?

 We cut operational water use by 20% from 2010 to 2014. In 2014, we audited four of our highest water use sites; two in India, and one in Italy, and Singapore, respectively.  We cut water use at these sites by repairing leaks, investing in efficient equipment, and introducing more water-efficient cleaning procedures.

	Water consumption is just one dimension of water impact. We also consider aspects such as water scarcity and local water quality, which includes the release of pharmaceuticals from our operations.  Site environmental, health, and safety (EHS) audits seek assurance on governance, management systems, and performance, which includes an assessment of wastewater management practices. Where performance gaps are identified, we work with suppliers on improvement programs. We monitor existing suppliers via our team of relationship managers and through our Responsible Procurement Audit program.

	More than 80% of the water used across our value chain is in producing raw materials, and much of this is from agricultural produce such as milk, sugars, and eggs. We partnered with TERI, a sustainable development non-governmental organization in India, to develop a water diagnostic tool to identify opportunities for suppliers to reduce their water impact. In 2014, we piloted the tool to explore ways to reduce water use for suppliers facing the highest water stress.  

How is GSK using advanced manufacturing technologies to reduce the environmental impact of manufacturing operations?

 A manufacturing technology roadmap was developed and approved by the GSK board in 2013 with the aim of enhancing product quality, while also making our medicines more accessible to patients anywhere in the world, in cheaper and greener ways. A team to seek, incubate, and commercialize advanced manufacturing technologies (AMT) was formed in 2014 to deliver this aim. Working with our product development and manufacturing organizations and across GSK, this team is enabling the application of advanced technologies to commercial processes.

	Four specific areas of the roadmap that have a significant impact in reducing the environmental impact of our processes are: synthetic biochemistry, chemical catalysis, continuous processing, and process analytics and control. Intensifying processes allows us to reduce solvent and material use, increase quality and yield of our products, and reduce the size of buildings and energy used in manufacturing.

	For example, our API manufacturing plant at Quality Road in Singapore replaced the chemical process used to manufacture the antibiotic, amoxicillin, with a process that uses a biology-based technology using enzymes.  The enzymatic process replaces organic solvents with water, so almost 80% of the chemical waste associated with older chemical processes has been removed. The new process helped us to reduce our amoxicillin value chain carbon footprint by up to 12%. This sustainable approach to the manufacture of amoxicillin is consistent with GSK’s commitment, in partnership with the Singapore Economic Development Board, to improve the efficiency of pharmaceutical and fine chemical manufacture in Singapore. In June 2015, we announced we would be investing an additional £38M (S$77M) in the Quality Road manufacturing plant in Singapore, building on its initial investment of S$60M committed in 2012

 What role do you expect continuous manufacturing to play in ‘greening’ pharma manufacturing?

 With continuous manufacturing, less is more. A reactor is much smaller but we can do it much more quickly, in smaller reactors based on flow through a pipe. This is designed to precisely control the chemistry so that by the time the chemicals leave the tube, the product is made. Because production is continuous, we don’t need to stop, cool, or clean the reactor, eliminating losses to the local environment and reducing waste. By using this approach, tiny flows of 1 to 200 mL per minute can be as productive as a 4000-L traditional reactor if operated 24 hours a day for 300 days a year.

	In October 2014, we announced an investment of $27M (S$38.5M) for a continuous manufacturing facility in Jurong, which produces one of GSK’s key APIs. With the new technology, a plant of only about 100 m

facility needed for current methods. In the future, we aim to apply this technology to allow us to make products to demand, allowing us to reduce inventory and waste in our supply chains.

:
	When referring to this article, please cite it as J. Markarian, "Reducing the Environmental Impact of Manufacturing," 

DAVID LEAHY/GETTY IMAGESThe pharmaceutical industry has an important role to play in implementing solutions to global envi

Reducing the Environmental Impact of Manufacturing

	The advantages of single use systems (SUS), including lower capital cost and increased flexibility, are well understood. Some disadvantages, however, have grown larger than life due to a lack of understanding and a low level of comfort with something new, says Chris Smalley, director of engineering in the Sterile & Validation Center of Excellence, Global Technical Operations at Merck. "Stainless steel and glass don't have to prove themselves, but single-use plastic is the 'new kid on the block'." Issues that are seen as obstacles include extractables and leachables, container integrity (i.e., leaks), and waste disposal. Various industry groups have been working to increase understanding of how to get past these obstacles and to develop standards to drive acceptance of SUS. For example, an updated version of the American Society of Mechanical Engineers (ASME) Bioprocessing Equipment Standard includes a section on shelf life and storage of SUS (1). The Bio-Process Systems Alliance (BPSA) published an update to its 

Single-Use Manufacturing Component Quality Test Matrices Guide

 in November 2015 (2). In addition, the International Society for Pharmaceutical Engineering (ISPE) is authoring a 

	The industry has been talking about standardization since 2009, and, although all the issues have not yet been resolved, knowledge has been developed and significant progress has been made, says Smalley. Collaboration between users and suppliers has been one of the keys to this progress, as users have more fully delineated their requirements and communicated with suppliers about how to meet these needs. 

	Clear user-requirement specifications are needed to mitigate the risk of technology selection and qualification, and a partnership between users and suppliers is needed to minimize risks throughout the lifecycle and to have better change control, said Sabrina Restrepo, associate director of component engineering in the Sterile & Validation Center of Excellence, Global Technical Operations at Merck, in a presentation at the ISPE annual meeting (3). "If you have a design space and clearly understand risk, you can better deal with changes because you can see how they might affect the process," she explained. 

	"Users should implement SUS with the same validation procedures they would use with a stainless steel system," added Smalley in a presentation (4). "First, define the user requirements, and identify specifications, such as processing temperature. Have a knowledge repository and a system for handling documentation." One small difference in SUS documentation is that some items in a data-set about the part might come from the supplier of the plastic tubing, for example, and other information might come from the supplier of the plastic material used to make the tubing. Users should also note that the same component may have different requirements depending on where in the process it is used, and any inventoried part should be identified as to where it is approved for use. For example, a bag used in buffer storage may have less stringent requirements for particulates, extractables, and leachables than the same bag used in fill/finish.

	Understanding the risk of extractables, which depend on the component material, and leachables, which depend on the actual use of the SUS in the pharmaceutical process, is crucial and requires collaboration between users and suppliers, explained Malik Belattar, deputy director of the nonprofit organization, the Single-Use Technology Assessment Program (SUTAP), based in Switzerland (5). SUTAP was founded in December 2014 for the purpose of facilitating single-use standards development.

	Users and suppliers are now cooperating on both the individual company level and the industry level. The BioPhorum Operations Group (BPOG), which is primarily an end-user group, and the BPSA, which is primarily a supplier group, are now doing some projects together. For example, in two ongoing BPOG-BPSA collaboration efforts regarding 

 and user requirements, data from surveys taken by participants have been leveraged to better understand the opportunities. These efforts should drive increased use of SUS by creating a more robust understanding of its capabilities. "Presenting a unified message to regulators will help to show that industry has gained understanding of SUS and will alleviate some of the fear of the newness of SUS," suggests Restrepo. The high level strategy was published last October (6), and authors are encouraging readers to provide feedback via 

		ASME, Bioprocessing Equipment Standard (New York, NY, 2014). 	

		BPSA, "Single-Use Manufacturing Component Quality Test Matrices Guide," 

http://www.bpsalliance.org/wp-content/uploads/2015/11/BPSA_Component_Quality_Test_Matrices_FINAL_November_2015.xlsx

		S. Restrepo, "Development of Standard Tools for Risk Assessment Evaluation Related to Single-Use Technologies," presentation at the ISPE Annual Meeting (Philadelphia, PA, 2015).	

		C. Smalley, "Single Use Systems-Why the New Kids on the Block Need to Prove Themselves," presentation at the ISPE Annual Meeting (Philadelphia, PA, 2015).	

		M. Belattar, "How to Make E&L from an Unidentified Risk to a Managed Risk,” presentation at the ISPE Annual Meeting (Philadelphia, PA, 2015).	

		T. White and K. Ott, BioProcess Intl. 13 (9) 24-29 (2015).

Collaboration between users and suppliers to increase understanding, clearly define user requirements, and mitigate risk is facilitating acceptance by industry and regulators.

Overcoming Challenges in Implementing Single-Use Systems

	Biosimilar development requires comparison with an originator’s molecule to establish biosimilarity. “If found to be ‘similar’ during extensive physical, chemical, and biological characterization, subsequent nonclinical and clinical studies to demonstrate the same safety and efficacy profiles as the originator may be reduced,” note Fiona Greer, global director at SGS Life Science Services, and Rabia Hidi, director for the Biomarkers and Biopharmaceutical Testing Unit at SGS in France. Characterization is also an aid to design the manufacturing process to deliver the target product and to determine the potential for determining biosimilarity interchangeability according to regulatory guidelines. Although FDA’s formal guidance for determining interchangeability has not yet been released, various analytical approaches may be part of this determination. 

 spoke with Greer and Hidi about some of the analytical approaches used to determine biosimilarity.  

: What are the common approaches to determine biosimilarity?

: Common strategies at this stage all include assessment of primary and higher-order protein structure to examine defined quality attributes. Many batches of originator and biosimilar are studied for variation, particularly throughout the timeline of the manufacture of the originator.

	In practice, analytical characterization will follow the requirements of the ICH Q6B guidelines (1), including determination of primary amino acid sequence; post-translational modifications, such as disulfide bridges and glycosylation; and spectroscopic profiles. Various chromatographic, spectroscopic, and electrophoretic methods can be used to interrogate and compare molecules on the basis of size, charge, and shape. Co- and post-translational modifications, fragmentation, aggregation, deamidation, and oxidation should all be studied and compared both qualitatively and quantitatively. It is important to assess the three-dimensional structure of the molecule, as changes here can greatly impact biological activity-both in terms of efficacy and potential immunogenicity. Once a robust analytical package of physicochemical tests has been performed, these are complemented by appropriate functional assays. The aim is to build a bridge between the observed analytical structural characteristics and their impact on clinical safety.

: What are some of the challenges with determining biosimilarity and how can these concerns be handled? 

: One of the challenges is in relating any observed small differences in structure during the comparability exercise to biological activity and clinical safety and effectiveness. For example, slight changes in the glycosylation pattern of certain therapeutic proteins can have a significant impact on the stability, clearance, binding, immunogenicity, and effector function. It is important, therefore, that appropriate 

 functional assays are performed. The type of functional tests should be based on the mechanism of action of the drug being studied. Ideally, bioassays should allow an assessment of all functional domains of a biosimilar candidate during comparison to the originator. Monoclonal antibodies are examples where multi-functionality should be assessed. Conventional assays for testing the functions of fragment antigen-binding (Fab) and fragment crystallizable (Fc) domains of therapeutic antibodies are widely available. These assays include 

 target binding, either on intact cells or using a soluble target; antibody-dependent cell-mediated cytotoxicity (ADCC); complement-dependent cytotoxicity (CDC); programmed cell death (PCD); and surface plasmon resonance (SPR) Fc-receptor binding assays.

: >What tests do you use to determine biosimilarity?

: Biological activity for comparability testing is performed using a suitable quantitative potency assay, which has been fit-for-purpose validated using the best experiment design. These assays should be developed to offer high precision and sensitivity for 

 detection of functional differences between the biosimilar and the reference compound. The design of the experiment is crucial during the development and the validation of these assays and should take into account the intended application of the method, such as the number of samples to be tested in one run and the number of replicates.      

	We have previously presented data of two experiments performed within our laboratories that examined functional activity of a biosimilar to its reference product (2, 3). In both these instances, a reference medicinal product is compared with a biosimilar for insulin. In the first set of data, we assay for 

 potency testing of insulin reference medicinal products and biosimilar compounds for inducing cell proliferation. Insulin and analogs are mediated predominantly via stimulation of the insulin receptor, but also through the insulin-like growth factor-1 receptor (IGF-1R). The stimulation of the IGF-1R and the effect on 

 mitogenic activity represents an indicator during 

 comparability testing. The second set of data demonstrates a receptor-binding assay undertaken to establish comparability in reactivity.

	Prior to analysis of comparability data, outlier values from the biosimilar test sample and the reference medicinal product are detected, using a statistical model chosen during the development and confirmed during the validation of the method. A four-parameter logistic (4PL), nonlinear regression model is commonly used for curve-fitting analysis in bioassays, immunoassays such as enzyme-linked immunosorbent assays, or dose-response curves. Then, the system suitability is assessed to check whether the preparation of the samples behaved in a manner consistent with the expected performance of the assay. This includes adequacy of the 4PL fit to be used to assess similarity (reflected by R

 values). Other system suitability criteria are the background and maximum responses, top/bottom of the curves (= specific response), slope of the curves, the EC50 or IC50 values (corresponding to the concentrations of the drug showing 50% of the maximum stimulation or inhibition response, respectively) and the variability between replicates of signal data. 

	To be used for biosimilarity comparison, the dilution curve from the biosimilar test sample must demonstrate parallelism when fitted against the dilution curve of the RMP. In other words, the two have sufficiently similar concentration–response curves or similar straight-line subsets of concentrations.  Parallelism of the two superimposed curves will be obvious by visual inspection of the plots and also confirmed by running an appropriate statistical test which should demonstrate that the linear parts of the curves are parallel.

	Once parallelism of the data is demonstrated, then the relative potency is determined using EC50 (or IC50) values determined from biosimilar and RMP dilution curves. Depending on the variability and the robustness of the method, it is recommended that potency results be reported from a number of repeats to ensure that the most accurate combined single potency value is conveyed.

Specifications: Test Procedures and Acceptance Criteria for Biotechnological/Biological Products

, CPMP/ICH/365/96 (September 1999).
	2. S. Roussi, et al., “Development and validation of a mitogenesis potency assay to be used for 

 potency testing of insulin biosimilar compounds,” Poster at the AAPS National Biotechnology Conference (San Diego, 2014).
	3. E. Stephan, et al., “Development and validation of a receptor binding competitive radioactive assay to be used for potency testing of human insulin binding to insulin receptor,” Poster at the AAPS National Biotechnology Conference (San Diego, 2014).

:
	When referring to this article, please cite it as J. Markarian, "Determining Biosimilarity," 

Analytical methods and functional assays are used to compare molecules and relate characteristics to quality attributes.

Determining Biosimilarity

	Next-generation pharmaceutical manufacturing-which must be more efficient, lower cost, and faster to market, while maintaining or improving quality-was a recurring theme at the International Society for Pharmaceutical Engineering (ISPE) annual meeting (Nov. 8–11, 2015, Philadelphia, PA). Although many discussions focused on the high-growth area of biopharmaceuticals, oral-solid dosage (OSD) is a large volume, globally important segment that is here to stay. The 2015 overall winner of the ISPE Facility of the Year Awards, in fact, is an OSD facility-AstraZeneca China's 533,000-ft

 solid-dose facility built in Taizhou, China for local supply of OSD drugs. 

	OSD drug manufacturing is mature and has been slow to change, but it is time for OSD manufacturing to embrace next-generation technologies, noted presenters from ISPE's OSD Community of Practice at the annual meeting. This group authored the third edition of the 

, which will be published in the first quarter of 2016 (1). The new edition considers new technologies available, such as process analytical technology for controls and instrumentation, and other changes in regulations and best practices, such as automation to reduce the risk of human contamination.

	A new chapter in the guide addresses containment and cross-contamination, which are increasingly important in OSD because of the growing use of highly potent APIs. It is important to identify parts of the OSD processing equipment that are crucial for containing potent materials, explained Richard Denk, sales for containment at SKAN, in a presentation. These include direct openings in the process where material transfer takes place, such as API dispensing, as well as equipment such as filters in the deduster, which may need containment when opened for cleaning and maintenance, noted Denk (2). Packaging equipment is another aspect to be considered for containment, because operations such as blister filling could potentially expose product.     

	Another area addressed by the guide is the growing regulatory pressure to produce OSD drugs in classified space, explained Norman Goldschmidt, vice-president at Genesis, Architects, Engineers and Constructors, in a presentation at the meeting (2). The US FDA doesn't require classification for OSD manufacturing spaces, but the China FDA (CFDA) is looking for Grade D classification, he explained. This is similar to the design standard of ISO-8 "at rest," which is suggested in the World Health Organization (WHO) TRS-961 Annex 5 (3), although Grade D has some additional testing requirements. Achieving this level is not burdensome, but can be achieved with air changes in the range of 5 to 20, said Goldschmidt. The new Baseline Guide does not recommend a specific air change rate, however, because the appropriate number depends on many factors, including the potency and form of the product and dustiness of the process step. Dilution of pharmaceutical powders via HVAC [heating, ventilation, and air conditioning] is not necessarily the best or only way to control contamination, cautioned Goldschmidt, who pointed out that the guide discusses various methods for environmental control and containment, including isolation, closed operations, and automation.

	Having a better understanding of new technologies and using them in manufacturing processes will help OSD manufacturers meet regulatory and cost pressures and become more efficient and effective.

		ISPE, Oral Solid Dosage Forms Baseline Guide, Volume 3 (Tampa, FL, in process).	

		Education Session, “The Latest OSD Regulation Requirements and Technologies,” presentations at the ISPE Annual Meeting (Philadelphia, PA, 2015).	

WHO Expert Committee on Specifications for Pharmaceutical Preparations, WHO Technical Report Series, No. 961, Annex 5 (Geneva, Switzerland, 2011).   

Experts at the ISPE annual meeting describe best practices, including containment and production in classified spaces.