Pressure BioSciences (PBIO) is developing a number of unique technology platforms involving the application of high hydrostatic pressure to accelerate and enable higher-quality research, drug discovery and development, and the manufacture of cannabinoid (e.g., CBD) products. David Alvaro, editor at Pharma’s Almanac, sat down with Pressure BioSciences Chairman and CEO, Richard T. Schumacher, to discuss these promising developments.

Making the Application of High Pressure Safe

“Pressure is an amazing force of nature,” asserts PBIO founder, President and CEO Richard T. Schumacher. “It moves at the speed of sound, has the power to exquisitely manipulate molecules, and can be turned on and off like a light switch,” he adds. These are tremendous advantages compared with conventional methods for controlling molecules in the laboratory, particularly the application of hot and cold temperatures on chemicals. While raising and lowering the temperature can speed up or slow down molecular action, once temperature has been applied, it takes time to bring the system back to equilibrium. Similarly, chemicals can also be used to manipulate molecular action; the chemicals are then often removed, which requires both time and cost. Pressure can be used to effectuate many of the same changes that temperature and chemicals cause, but pressure can be instantaneously applied and removed, often with fewer negative consequences.

The challenge has always been to develop high-pressure processes that are safe yet effective in the laboratory setting. High pressure is currently used in various manufacturing sectors, but it is more challenging — and expensive — to design safe laboratory equipment that can go to tens of thousands of pounds per square inch (psi). As a result, research scientists have generally been limited to the use of pressures of a few hundred to approximately 10,000 to 12,000 psi.

PBIO has developed pressure-generating instruments that can apply pressure up to 45,000 psi (about three times the pressure experienced six miles below sea level at the bottom of the Marianas trench) in a safe, reproducible, and routine manner. The system employs hydrostatic (water) pressure; water is safely compressed in the instrument’s pressure chamber to high and very high levels; this pressure is then transferred to samples (e.g., normal cells, cancer cells, bacteria and viruses, biopsy tissues) contained in specially designed, consumable reaction vials in the pressure chamber. A next-generation instrument under development can reach pressures as high as 100,000 psi, according to Mr. Schumacher. PBIO has 24 patents on its high-pressure based Pressure Cycling Technology (PCT), BaroShear Technology (BaroShear), and Ultra Shear Technology (UST^TM) platforms.

New Weapon for Increasing the Quality of Research Studies

In most of PBIO’s instruments, pressure can be turned on and off repeatedly, which is referred to by the company as PCT (pressure cycling technology). Of the company’s 24 issued pressure-based patents, over half relate to the PCT methodology.

One of the widest applications for PCT is in the investigation of cellular materials, such as proteins, DNA, RNA, and lipids. By repeatedly squeezing cells with hydrostatic pressure (akin to squeezing a sponge), PBIO’s GMP-compliant, next-generation PCT-based Barocycler EXTREME instrument enables the controlled release of cellular material, which can then be used for the design, development, characterization, and quality control of biotherapeutic drugs; in the discovery of biomarkers for such important medical conditions as cancer, heart disease, and Alzheimer's; and in forensics.

An advantage of using PCT is higher quality. The proteins and other materials extracted from cells broken with PCT are more likely to be in their natural state than traditional techniques for achieving cell lysis –– spinning with beads in test tubes or chopping with blades. PCT can further allow access to proteins and enzymes not seen when cells are mechanically or chemically degraded; 5–10% of the total proteins obtained using PCT are only observed when this method is used. “Our PCT platform provides scientists access to a larger number of potential drug targets, biomarkers, and other important discoveries than traditional sample preparation methods,” Mr. Schumacher states.

The PCT technique is also accelerating the investigation of proteins once they are isolated from cells. Protein digestion (breaking the protein down to its building blocks) is a critical part of scientific research. Using conventional techniques, protein digestion often takes hours and can be difficult to perform under consistent conditions. Using PCT, protein digestion is generally achieved in minutes and is completed under controlled conditions, in the same manner, every time, assuring that the sampling and processing conditions do not influence the results. The company currently has over 300 PCT instrument systems placed in approximately 225 academic, government, pharmaceutical, and biotech research laboratories worldwide.

Addressing Folding and Aggregation Issues

Misfolding (undesired folding configurations) and aggregation (clumping) of proteins being developed for potential therapeutic purposes directly impacts their efficacy and can also have serious safety consequences. PBIO has discovered that some proteins with incorrect structural configurations, when placed under pressure, under certain conditions, will denature and subsequently return to their proper, natural configuration upon removal of the pressure. The result is that the protein can now exert its intended purpose. For a protein being developed to fight a disease, fix a disorder, or prevent infection, this fix could be crucial.

The company’s BaroFold Platform employs high pressure for the disaggregation and controlled refolding of proteins to their native structures at yields and efficiencies not achievable using existing technologies. It has been shown to remove protein aggregates in biotherapeutic drug manufacturing, thereby improving product efficacy and safety for both new-drug entities and biosimilar products.

PBIO currently has three clients for which PBIO is determining whether high-pressure application can address structural issues and are seeking more partnerships. Some of these clients will purchase the Barford instrument and conduct the initial investigations themselves. Others will have PBIO do the initial testing. For any potential drug that benefits from the technology, PBIO will then support the sponsor firm through the IND development stage, approval process and every commercial manufacturing batch throughout the life of the product. The company believes this could result in millions of dollars in services performed and in-use licenses for years to come.

Nanoemulsion Technology to Overcome the Insolubility of CBD and Other Cannabinoids in Water

PBIO’s third platform, Ultra Shear Technology™ (UST™), is based on the use of ultra-high pressure combined with intense shear forces. A liquid material (e.g., CBD oil, milk, skin product or pharmaceuticals) is placed in the company’s specially designed, custom-made, high-pressure–based instrument (the BaroShear), subjected to ultra-high pressure, and discharged at high pressure through a special flow-path that leads to a patented dynamic (nano-gap) valve under ultra-high shear conditions, destroying most (if not all) pathogens in the liquid, with the resulting formation of a highly stable, water-soluble nanoemulsion.

UST has been shown to turn hydrophobic (water-repelling) extracts into stable, water-soluble formulations on a small, laboratory-scale and offers the potential to produce stable nanoemulsions of oil-like products in water, with applications in many markets, ranging from food and beverages to inks, paints, and cosmetics, as well as in pharmaceuticals and nutraceuticals, such as medically important plant oil extracts.

The company is initially focusing its UST platform in two areas. First, through a partnership with Ohio State University, and funded with a four-year, $1 million grant from the U.S. Department of Agriculture (USDA), a system is being developed and patented that will enable the production of small and large amounts of milk and other dairy products that are shelf-stable for months at room temperature.

A second project involves the production of cannabidiol (CBD) nanoemulsions. CBD and other cannabinoid derivatives are isolated from the hemp plant as a poorly water-soluble oil. As a result, only about 10% of the CBD is extracted by the body before the CBD oil is excreted. Numerous companies are exploring chemical modification and encapsulation methods for improving the solubility of CBD, according to Mr. Schumacher. UST, on the other hand, simply (but not trivially) increases the surface of the area of the CBD oil by shearing large oil drops containing millions of CBD molecules into millions of nano-sized droplets containing only a few CBD molecules, all of which remain unchanged and can be more effectively absorbed by the body.

The BaroShear K45 System (for which presales were recently announced) has a maximum throughput capacity of approximately 1 liter per hour and is suitable for processing small (20 mL minimum) to moderate volumes of high-value CBD oil to nanoemulsion levels with minimal loss during production. PBIO is developing a small, benchtop version of the BaroShear instrument for small-volume processing and R&D activities, as well as a much larger version for very high–volume production. PBIO is also developing the UST Platform for commer­cialization, with an expectation that initial orders for the BaroShear K45 will be installed and billed by late 2020.

Small Company Tackling Big Issues

The biggest challenge for Mr. Schumacher today is the fact that PBIO is a small company with limited funding and a vast array of opportunities, only a few of which the company can pursue at present. “If we were a larger, well-funded company, we could develop multiple product and service lines around each of our technology platforms targeting multiple applications, said Mr. Schumacher. For the moment, PBIO is focusing on the PCT platform in cancer, the BaroFold platform for protein therapeutic development, and the UST platform for CBD nanoemulsions. “By pursuing these three key initial opportunities, we believe we will, in the near future, finance PBIO properly and be in a position to explore the endless opportunities that these technologies provide,” he concludes.