NMR: A zooming lens for Biopharmaceutics

Work done in the lab of Prof. Ashutosh Kumar at Department of Biosciences and Bioengineering, Indian Institute of Technology Bombay.

About author

Soumya Ranjan Pujahari, a doctoral candidate at the Department of Biosciences and Bioengineering at IIT Bombay, is currently being supervised by Prof. Ashutosh Kumar and co-supervised by Dr. Navratna Vajpai from Biocon Biologics Ltd. His educational background includes a bachelor’s degree in Biotechnology from the Central University of Rajasthan and a master’s degree in biotechnology from the University of Hyderabad. His research primarily focuses on the analytical characterization of biopharmaceutical products, with several of his projects being carried out in collaboration with Biocon Biologics Ltd., located in Bangalore.

Interview

How would you explain your research outcomes to the non-scientific community?

Biopharmaceutical drugs encompass medicines sourced from living organisms or created through biotechnology. They offer precise and often intricate therapeutic approaches to treat diverse diseases. These drugs fall into two primary categories: biologics (also known as reference drugs) and biosimilars. Biologics are groundbreaking and intricate medications, protected by a 20-year patent. In contrast, biosimilars are highly similar replicas of approved biologics, exhibiting no clinically significant disparities in safety and effectiveness. Biosimilars enter the market only after the expiration of the patent duration for the biologics. The significance of biosimilars lies in their ability to enhance patient outcomes and decrease healthcare expenses. By providing more affordable alternatives to costly biologic drugs, biosimilars enhance access to treatment for patients dealing with chronic and life-threatening conditions like cancer, rheumatoid arthritis, and diabetes. This increased accessibility significantly impacts a patient’s quality of life and overall health.

However, it is crucial to acknowledge that, despite their resemblance to reference biologics, biosimilars are not exact copies. Stringent regulatory procedures guarantee their safety, effectiveness, and quality, subjecting them to extensive analytical and clinical testing to establish their similarity to the reference product. In accordance with regulatory guidelines, biosimilar products must closely resemble marketed products in all aspects, including the establishment of similarity in higher-order structures. Various analytical methods are employed to demonstrate the similarity in higher-order structures between reference products and biosimilar drugs. It is particularly challenging to characterize the biopharmaceutics of a drug in its natural state, and this difficulty is amplified in suspension formulations where both solution and solid phases coexist (known as biphasic drugs). Nuclear Magnetic Resonance (NMR) is an advanced biophysical technique capable of providing high-resolution structural information about a biologic drug in its formulated state. In our study, we have developed an NMR-based approach to demonstrate the similarity between two biphasic drugs without disturbing their original state. The obtained data is then subjected to analysis using various statistical tools to quantitatively evaluate the similarity.

A higher-order structure (HOS) is critical to a biopharmaceutical drug as the three-dimensional structure governs its function. Here, we have used a combinatorial approach using liquid (1D 1H) and solid-state (13C CP-MAS) NMR methodology to demonstrate the HOS in the biphasic microcrystalline suspension drug in its formulated state. The data were further assessed by principal component analysis and Mahalanobis distance (DM) calculation for quantitative assessment.

How do these findings contribute to your research area?

The results of our research will have a direct positive impact on the biopharmaceutical sector, particularly in India. Currently, India holds a prominent position as a leading exporter of biopharmaceuticals, with a focus on biosimilar products. The development of biosimilar products involves rigorous biophysical characterization to meet regulatory standards. Our approach can significantly contribute to demonstrating the similarity between two microcrystalline products, a crucial factor in biosimilarity evaluation. Furthermore, our method can be beneficial in the initial stages of product development, during storage, and in monitoring batch-to-batch variation.

“Our approach can significantly contribute to demonstrating the similarity between two microcrystalline products, a crucial factor in biosimilarity evaluation.”

What was the exciting moment during your research?

While developing this technique, we discovered intriguing findings and formulated various hypotheses. To validate our hypotheses, we employed several different approaches and orthogonal techniques, and remarkably, each outcome supported our proposed statements. This outcome was truly gratifying.

What do you hope to do next?

We are consistently making efforts to enhance our understanding of biopharmaceutical products by exploring various analytical methods. In the coming years, we aim to expand upon our current approach and make further improvements to it.

Where do you seek scientific inspiration from?

I derive my scientific inspiration from my Ph.D. mentors, Prof. Ashutosh Kumar, and Dr. Navratna Vajpai. I feel the need to highlight both individuals because whenever I encounter difficulties, I simply inform them, and they are consistently available for scientific discussions, regardless of the day or time. In addition to them, Dr. A.P.J. Abdul Kalam serves as both a scientific mentor and a role model for me.

How do you intend to help Indian science improve?

Given my current involvement in the biopharmaceutical sector, I desire to utilize and apply my expertise to enhance the Indian biopharma industry. Additionally, I aim to contribute as a liaison connecting academia and the pharmaceutical sector.

Reference

Soumya Ranjan Pujahari, Pramod S. Mali, Rudra N. Purusottam, and Ashutosh Kumar*. Combined Liquid-State and Solid-State Nuclear Magnetic Resonance at Natural Abundance for Comparative Higher Order Structure Assessment in the Formulated-State of Biphasic Biopharmaceutics. ACS Analytical Chemistry, 2023

https://doi.org/10.1021/acs.analchem.2c05485

Copy Editor

Sukanya Madhwal

Ph.D. student at inStem Bangalore

Sukanya hails from a small town, kotdwara in Uttarakhand. She completed her Masters’ degree in Biotechnology from Banasthali Vidyapith, Rajasthan. After this, she served as a graduate teacher for one year at P.G. College, Kotdwara, Uttarakhand. In 2014, she qualified for the JGEEBILS exam conducted by NCBS/TIFR and joined as a Research Scholar (Ph.D. student) in Dr. Tina Mukherjee’s lab at inStem. Currently, she continued working as a bridging post-doc in Dr. Tina Mukherjee’s lab. Besides work, she loves reading non-fiction books, enjoys gardening, and cooking delicious food.

For interview related queries, write to us at interview.biopatrika@gmail.com