Biosimilars: Expanding Manufacturing and Regulatory Horizon for Creating High Value Market
Dr Madhusudan P Dabhole, Group Manager- Bioprocess, Richcore Life Sciences Ltd.

Biosimilar industry has been growing stupendously. In the last few years, India has seen a robust growth in its biosimilar portfolio. This article aims to identify and analyse biosimilar market in India.

Biological research and biologics has perceived a quantum upsurge in all areas from thinking about making a molecule in laboratory to commercial aspect of marketing and response. The Biopharmaceutical industry has gained a lot of impetus and has shown a commendable progress. Starting from Recombinant Human Insulin which was the very first therapeutic biopharmaceutical product to be approved to Antisense technology used today as a drug discovery platform, biopharmaceutical has come a long way. With the expiration of patents of innovator biologics, biosimilars have created opportunities for this industry to mature. Due to their high degree of specificity, biologics are the drivers of growth in the pharmaceutical industry in the long term.

"The identification of double helix has been a landmark discovery in the history of science and I am very proud to be associated with it", said Dr James D Watson, sitting outside the auditorium at Tata Institute of Fundamental Research (TIFR) Mumbai, after finishing his lecture on Genes and Politics on December 1st 1997.

I wondered why Dr Watson had not acknowledged the contribution of Rosalind Franklin and posed the question to Dr Watson. He smiled and said,"I am not selfish". When asked about the significance of sequencing the human genome and its benefits in understanding disorders, he added, "Now that the Human Genome is being sequenced, proteins will be the key molecular machines which will play an integral part in future". I was searching something distinct on which I can take his autograph and found one small Lord Ganesh greeting. I moved forward and took his autograph which he signed off. I decided to pursue my PhD in Stress protein application.

Biosimilar is a biopharmaceutical drug designed to have active properties similar to one that has previously been licensed. The USFDA defines Biological products as a wide range of products such as vaccines, blood and blood components, allergenics, somatic cells, gene therapy, tissues, and recombinant therapeutic proteins. Biological can be composed of sugars, proteins, or nucleic acids or complex combinations of these substances, or may be living entities such as cells and tissues.

Regulatory Hurdles in Biologics
The Food and Drug Administration's (FDA) Center for Biologics Evaluation and Research (CBER) plays a leading role in the development of therapies that will become the standard of care in the 21st Century, both in the United States and around the world. These emerging therapies include a wide variety of biologic products, such as those for gene replacement and tissue regeneration. In addition, novel strategies and approaches may be employed for developing new biological products.(6)

CBER-regulated products pose four significant regulatory challenges that reflect both the rapid pace of biomedical research and the nature of the products the center regulates:
  1. Complexity and diversity: Biologics are complex and diverse, and often incorporate new scientific knowledge. CBER reviewers must have an in-depth understanding of the technology used to prepare them, and when feasible, knowledge of the mechanism of action.
  2. Vulnerability to contamination: Biologic source materials used to manufacture products (eg, mammalian cell lines) and the source of the product itself (eg, blood from human donors) are particularly vulnerable to contamination with infectious agents.
  3. Processing difficulties: Biologics cannot withstand common purification and decontamination methods, and thus require rigorous control of the source materials, manufacturing processes, and sensitive and specific safety, potency, and purity testing.
  4. Long-term adverse effects of treatment: Some biological products, for example certain infused cells or integrating viral vectors, have the potential to persist long-term in the body. Some types of adverse events , which might include cancer, may take years to occur, and understanding how to monitor and clinically respond to such adverse events poses unique challenges.(6, 7)
Biologics and Clinical Trials
In the last decade, India was considered as one of the major players in the clinical trials field due to the relatively low costs of R & D in India. But in the recent past, it was observed that there is a need for a medical ethics committee which has been inducted to control the clinical trials under the lens minutely.

Another barrier is the difficulty in attracting patients to clinical trials of biosimilars. Patients may be reluctant to participate in the trials, especially for serious diseases, because only some of them will receive the biosimilar rather than being confident of receiving the branded biologic outside of a clinical trial.

Because many companies may be attempting to develop the same biosimilar, it may be difficult to get enough volunteers because they are competing for the same limited population. Based on discussions with people in the pharmaceutical industry, some biosimilar companies have encountered difficulties in obtaining the reference product for the trials, and when they do, it is often quite expensive.(8)

Rising cost in Clinical Trials involving the number of studies and subjects at different time intervals is another factor which needs attention. The pharma sponsors use multiple clinical research organisations (CROs) - a mix of full service and niche CROs. They also prefer to use functional CROs in outsourcing. This means that the whole clinical trial process may be shared by multiple CROs managing diverse functions - regulatory approval, protocol preparation, site management, monitoring, data management, statistics, and medical writing. This could mean multiple CROs using multiple standard operating procedures (SOPs) or the sponsor asking each one to follow the sponsor's SOPs.

The fragmentation of outsourcing can lead to deficits in documentation, unclear division of responsibilities between CROs and sponsors, and limited real-time assessment of the CRO function. It will also be difficult to carry out a comprehensive root cause analysis and corrective actions. The FDA is concerned that many third parties involved in the clinical trials can impact data integrity and/or human subject protection.(9)

Some common deficiencies that were observed and are being taken care during Quality of Clinical trial site inspections include:
  • Failure to follow the investigational plan and signed investigator statement/agreement.
  • Protocol deviations.
  • Inadequate record keeping.
  • Inadequate accountability for the investigational product.
  • Inadequate subject protection, including informed consent issues.
  • Adverse Event (AE) recording and reporting.
Currently estimated at USD 500 million, India's clinical research market was projected to more than double and cross USD one billion mark by 2016 driven by favorable factors like diverse and accessible population, availability of low cost and effective resources.(10)

Challenges in Developing Biosimilars Many biochemical and biophysical studies of proteins depend on the availability of milligram quantities of highly purified proteins of interest(3). Increasing development costs and a demand for more compelling demonstrations of the value of new products from regulatory agencies has increased the challenges for biologics development. The scale-up of a cell culture process can be very difficult and time -consuming, taking as long as several months before researchers can obtain a product.

The entire process of producing a biotech product from start to finish is often called a campaign and is usually divided into two main parts: upstream and downstream. Upstream processes involve production of the protein product , most often by using cells (microbial, insect or mammalian) growing in culture. Downstream processes include the recovery, purification, formulation and packaging of the protein product.

Variability is the killer. Whether running 150 m3 industrial processes or starting off a PhD with a couple of 'control' runs. Until the variability in any process is understood the results and any conclusions drawn will be open to interpretation, scrutiny and worst of all, error. Somewhat worryingly, variation not only applies to existing processes, but to experiments not even thought of! Hence, Use of statistical tools will enable interpretation of data and phrasing of conclusions. The statistical software widely used is Minitab. Sources of variability include, but are not limited to: raw materials, operator error, engineering/mechanical faults, documentation or calibration.

Control charts, Process capability, Regression, correlation and Fractional factorial design are some of the measurable tools that can be used for identifying and controlling variation.(5)

Microorganisms in nature are subject to both favorable and unfavorable influences due to varying chemical and physical environmental factors. Living cells like bacteria and yeasts, therefore, display a rapid molecular response when exposed to adverse environmental conditions. This is known as stress response and can be induced by various means such as exposure of cells to high temperature, low temperature, ethanol, hydrogen peroxide, sodium chloride, cadmium and mercury which induce stress proteins. To combat these metals, cells use numerous mechanisms such as solubilisation and metal uptake from the extracellular environment.

Various types of bacteria, filamentous fungi and algae have been used for metal removal from aquatic systems. Furthermore, there is very little information available about the uptake of cadmium and mercury by Rhodotorula mucilaginosa.(2)

In order for proteins to be used as pharmaceuticals, delivery technologies need to be developed to overcome biochemical and anatomical barriers to protein drug transport, to protect proteins from systemic degradation, and to target the drug action to specific sites. Protein transduction domains (PTDs) are used for the non-specific transduction of bio-active cargo, such as proteins, genes, and particles, through cellular membranes to overcome biological barriers.

Metallothionein (MT) is a low molecular weight intra-cellular protein that consists of 61 amino acids, including 20 cysteine residues, and is over -expressed under stressful conditions. Although MT has the potential to improve the viability of islet cells and cardiomyocytes by inhibiting diabetic-induced apoptosis and by removing reactive oxygen species (ROS), and thereby prevent or reduce diabetes and diabetic complications, all MT applications have been made for gene therapy or under induced over -expression of endogenous MT.

To overcome the drawbacks of ineffective intra-cellular MT protein uptake, a human MT gene was cloned and fused with protein transduction domains (PTDs), such as HIV-1 Tat and undeca-arginine, in a bacterial expression vector to produce PTD–MT fusion proteins.(17)

Logistics Issues
Cold Chain Management refers to the chain of logistics that keeps an item refrigerated from the time it leaves the factory until its final destination . Temperature Monitoring with Validation at all stages during Shipping, distribution and warehousing are major issues impacting Cold Chain Management.

The supply chain for biosimilars will be very different to the current range of generic drugs. Biopharmaceuticals are less stable than chemical based pharmaceuticals and thus require cold chain distribution and have a shorter shelf life. This increases the cost and complexity of distribution(1). Biopharmaceuticals are highly temperature sensitive and are recommended to be stored and transported at temperatures ranging from 2°- 8°C. Any failures in the cold chain transportation of these drugs can lead to a significant loss of their efficacy and can even make them harmful for the patient.

Regulatory agencies have stated that drugs that have been temperature mistreated are every bit as dangerous as those that are not authentic. The demand for cold chain logistic services is currently experiencing explosive growth. IMARC Group, one of the world's leading research and advisory firms, finds that the total size of the healthcare cold chain logistic services market is expected to expand from its current figures of USD 7.3 billion to nearly USD11.4 billion by 2018.

The Biopharma companies along with the Cold Chain Logistics services should address how to monitor and control temperatures with other associated risks to biopharma products from manufacturing across the whole supply chain to the end user.

Enhancing Cost Efficiency in the Manufacturing of Biologics
The inherent risk in establishing Biologics production (product, process, timeline, capacity, regulatory and location) can be significantly mitigated by using a modular and standardised approach. Utilising a combination of standardisation, modularisation and use of modern process solutions such as single use equipment offers significant advantages over traditional design and construction.(11)

The cost and complexity of developing and manufacturing biosimilars also suggests that, realistically, product pricing reductions will be 20 to 30 percent, compared to the 80 per cent typical for small molecule drugs. These relatively low margins, combined with pressure from healthcare systems on end-user pricing, will require less costly manufacturing alternatives than the big tank approaches now widely in use.

The need for multiproduct facilities, with increased flexibility, reduces the requirements for expensive critical utilities such as water for injection and clean steam, decrease requirements for equipment cleaning and cleaning validation, lower capital investments, and shorten facility construction times.

All of these realities have caused biopharma manufacturers to incorporate disposable or single-use process technologies into their product manufacturing. These systems offer a smaller footprint, flexibility, scalability, and mobility without compromising product quality.(12)

One key advantage of modular construction for biopharmaceutical facilities is the off-site construction of modules. The benefits of this approach include enhanced quality control, reduced waste, reduced impact on current operations, and simplified site logistics. Transferring labor hours away from the construction site also reduces risk and overall cost for a facility construction project.

Building multiple modular elements in parallel without, for example, weather impact, can reduce the construction schedule for a facility project by 50 percent. The ability to leverage factory acceptance testing (FAT) at a module construction facility will often significantly shorten the time for start-up and commissioning of a new facility. Once modules are delivered to the construction site, they are assembled into the complete facility so that final testing and qualification can be completed.(11)

Anti-Sense Technology in Drug Discovery
Antisense compounds are biological molecules consisting of small ribonucleic acid (RNA) or deoxyribonucleic acid (DNA) segments which have enormous potential for the treatment of number of diseases. The first report of usage of this particular antisense oligodeoxynucleotides to inhibit Rous sarcoma virus gene expression, there has been tremendous progress in the understanding and application of antisense oligodeoxynucleotides.

This antisense approach should allow the design of drugs that specifically intervene with the expression of any gene whose sequence is known by that it will be more convenient for the treatment for genetic disorders or infections Antisense drugs are being researched to treat a variety of diseases such as cancers including lung cancer, colorectal carcinoma, pancreatic carcinoma, malignant glioma and malignant melanoma, diabetes, amyotrophic lateral sclerosis (ALS), Duchenne muscular dystrophy and some other diseases such as asthma, arthritis and pouchitis.

The concept behind antisense technology is quite straightforward: the use of a sequence, complementary by virtue of Watson-Crick base pairs hybridisation . This Oligonucleotide is modulated to a specific mRNA which can inhibit its expression and then induce a blockade in the transfer of genetic information from DNA to protein, but mechanism of induction of its biological effect is subtle and complex. The aims for optimal delivery of antisense compounds are therefore to enhance cellular uptake and improved to exit from subcellular compartments as correct targeting (spatial and temporal) to a particular site of action. Liposomes are considered very promising delivery systems for antisense therapeutic approach, offering drug protection and facilitating oligonucleotide cell internalisation.(13)

Comparison of the Experimental Set up
  • Measurement principle of the microtiter plate fermentation in the BioLector via back scattering of light from cells and fluorescence emission of molecules;
  • Measurement principle of the stirred tank fermentation with online measurement of OD and fluorescence in the bypass.(4) Kensy et al. Microbial Cell Factories 2009 8:68.
New Forms of Drug Delivery
The practice of drug delivery has changed dramatically in the last few decades and even greater changes are anticipated in the near future. Clinicians historically have attempted to direct their interventions to areas of disease or areas at risk for disease. Depending on the medication, the way it is delivered, and how our bodies respond, side effects sometimes occur.

Microneedle arrays are one example of a new method to deliver medications through the skin. In these arrays, dozens of microscopic needles, each far thinner than a strand of hair, can be coated or filled with a medicine. The needles are so small that, although they penetrate the skin, they don't reach nerves in the skin, thus delivering medications painlessly.

Nanosponges created by NIBIB-funded researchers, are a promising vehicle in treating cancer. Comprising a scaffold of tiny, specialised polyester particles coated with disease-targeting compounds and filled with an anticancer drug, the nanosponges home in on tumors after being injected into the body. Once at their intended site, they safely and slowly degrade, releasing medication at the tumor site at a steady, controlled rate. Early studies have also shown the nanosponges can be used to treat glaucoma, the fourth leading cause of blindness.(15)

Corporate alliance deals structured as licensing transactions, Co -development agreements, joint ventures, or sales and marketing alliances play an integral role in many growth strategies for biopharmaceutical companies. Strategic partnerships can involve research focused on discoveries characterised by unique mechanisms of action at the molecular level and within cellular processes that can be targeted as a way to develop best-in-class medicines.(14)

The explosive growth of biologic medicine and the emergence of biosimilars as revolutionary tools to fight the most difficult of disease are to provide advanced health care to patients worldwide.

The future of biological medicines will be bright if patients, physicians, biotechnology companies, and other stakeholders work together to ensure patient safety which are the foremost priority of the biosimilar policy discussion. Then, the future of healthcare debate can move beyond parochial discussion over healthcare access and cost, to a discussion of the diseases that biological medicines can successfully conquer next.(16)

  1. Jo Pisani and Yann Bonduelle, Pharmaceutical strategy consultants at PriceWaterhouseCooper LLP: Opportunities and barriers in the biosimilar market: Evolution or Revolution for Generics companies?
  2. Madhusudan P Dabhole and Dr K N Joishy, Use of rapid indirect Elisa to detect cadmium bound to stress protein in Rhodotorula mucilaginosa, Ph.D Thesis, 2002.
  3. Uwe Riek et al, An automated home-built low-cost fermenter suitable for large-scale bacterial expression of proteins in Escherichia col. BioTechniques, Vol. 45, No. 2, August 2008, pp. 187–189.
  4. Frank Kensy et al - Scale-up from Microtiter plate to laboratory fermenter: evaluation by online monitoring techniques of growth and protein expression in Escherichia coli and Hansenula polymorpha fermentations Microbial cell factories: 2009, 8:68
  5. Brian Mc Neil and Linda M Harvey. Practical Fermentation Technology, John Wiley & Sons Ltd.
  6. Karen Midthun, Centre for Biologics Evaluation and Research, Strategic plan for Regulatory Science and Research, Draft - 2012-2016.
  7. Dawn Willow: The Regulation of Biologic Medicine: Innovator’s rights and access to Healthcare, Chicago-Kent Journal of Intellectual Property, 2006.
  8. Erwin A. Blackstone and Joseph P Fuhr, Jr , The Economics of Biosimilars, September/October 2013, Vol 6, No 8 - Business.
  9. Arun bhatt, Quality of clinical trials: A moving target Perspect Clin Res. 2011 Oct-Dec;2(4): 124–128.
  10. Priyesh Sharma, Vaish associates advocates, Future of clinical trials in India. Mondag, 28thJan 2013.
  11. Jan Lilja et al, Modularization in Biologics manufacturing. Pharma Focus Asia.
  12. Patricia Fitzpatrick Dimond, Flexibility in Biopharmaceutical Manufacturing Capacity. Single-use bioreactors are changing the biomolecular production landscape. Genetic Engineering and Biotechnology News, 2013.
  13. Lavakumar et al, Antisense technology: Oligonucleotides and its delivery strategy. British Biomedical Journal. 2014.
  14. Wendy Tsai et al, Early stage Biotech companies: strategies for survival and growth. Biotechnology Healthcare. 3(3), 49-50, 52-53, 2006
  15. National Institute of health. US department of Health. Drug Delivery Systems: Getting Drugs to their Targets in a controlled manner.
  16. Richard O Dolinar et al, The future of biological therapy: a pathway forward for biosimilars, Generics and Biosimilars Initiative Journal (GaBI Journal). 2013; 2(1):36-40.
  17. Yong hee Kim, Preparation and functional analysis of recombinant protein transduction domain-metallothionein fusion proteins, Biochimie, 92, 8,964-970, 2010.