Process Development
High-Throughput Biopharmaceutical
Drug Development
Meeting the Coming Challenge
he research component of
pharmaceutical research and
development (R&D) has been
undergoing a dramatic revolution in
the past five years, especially in how
promising new drug candidates are
identified. Throughout the past fifty years,
the pharmaceutical industry focused its
efforts on about 500 drug targets. Targets
are expected to number in the thousands
soon because combinatorial chemistry and
ultrahigh-throughput screening (up to
500,000 compounds per day) continue to
transform drug discovery efforts.
Using state-of-the-art technologies,
research groups can now deliver a hundredfold more compounds (in the same amount
of time) that warrant further investigation.
The Human Genome Project has created
new therapeutic avenues, offering the
potential for individualized medicine in the
21st century. Elucidating the complex
interrelationships of the approximately
30,000 human genes should lead to growth
in protein-based therapeutics for the near
future. Advances in genomics, proteomics,
bioinformatics, and pharmacogenetics are
accelerating the identification of protein
drug candidates tremendously (1).
T
Rajiv Nayar and
Mark C. Manning
Outsourcing is often considered a
way to expedite drug
development, but other options
exist for companies that don’t
choose it — or that run up against
the capacity shortage. The
resources devoted to speeding up
the drug discovery process led to
combinatorial libraries, highthroughput screening, proteomics,
and genomics. Now the same
types of innovation can be applied
to drug development to prevent
valuable lead compounds from
sitting idle on the shelf.
A Revolution in Research
Corresponding author Rajiv Nayar is president of
HTD BioSystems Inc., 551-C Linus Pauling Drive,
Hercules, CA 94547, 510.367.0528, fax
509.267.1491, [email protected],
www.htdcorp.com; and Mark C. Manning is chief
technical officer for HTD Biosystems and associate
professor of pharmaceutics at the University of
Colorado Health Sciences Center.
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BioPharm
FEBRUARY 2002
The shifting demographic profile of aging
baby boomers is stimulating the demand for
new medicines. Conditions such as
osteoporosis, arthritis, and dementia are in
the forefront, joining cardiovascular
diseases, cancer, and metabolic diseases.
Lifestyle expectations have led to a search
for better drugs to treat problems such as
obesity, acne, and erectile dysfunction.
Spurred by the growing demand for new
and innovative medicines, drug companies
are spending record amounts on R&D. In the
United States alone, R&D investments were
$26.4 billion in 2000, representing an almost
10% increase over 1999. The spending
estimate for 2001 is $30.5 billion, another
18.5% increase. Investment in
biotechnology alone was about $8 billion
(2). As a result of the incredible advances in
drug discovery, pharmaceutical pipelines are
filled with more products than ever. More
than 1,000 drugs are currently in
development, and nearly 350 of those are
biotechnology-related products (2). Table 1
breaks down the drugs in development by
disease indication.
A Crisis in Development
Researchers looking for new candidates are
finding many active compounds that merit
further investigation and development. The
result is that all development groups have
more projects going at one time than ever,
and that number will continue to rise.
Undoubtedly, an enhanced ability to develop
and formulate those drugs will be needed to
keep pace. In addition, increased use of
sophisticated drug delivery systems will
make stabilizing biotechnology-based
products even more challenging.
In short, the pharmaceutical
biotechnology industry must find new ways
to speed up drug development to keep pace
with drug discovery. We believe that the
limiting factor for introducing new
biotechnology-derived products is no longer
the search for new drug candidates but the
selection of those with the highest potential
for commercialization. Therefore, efficient
and rapid developmental strategies need to
be devised that cover all aspects of drug
development (such as fermentation,
purification, characterization, and
formulation).
The incentive for efficiency. Another driving
force for making drug development more
efficient is the pressure to contain health
care costs. Currently, the costs of R&D are
estimated at between $350–500 million for
Process Development
each drug approved (3). More than 90% of
the new drugs anticipated are predicted to
generate only about $180 million a year,
which is lower than the current industry
average of $265 million a year (3). With the
industry forecasted to exhibit a 7% annual
growth in sales (and assuming an increase of
7% in R&D spending), pharmaceutical
companies need to slash their R&D costs to
about $280 million for each drug approved.
That is the incentive for making drug
development, the costlier component of
R&D, more efficient. The current success
rate for development projects (1 in 10) is
unacceptable and financially unsustainable.
Developers should consider the situation as
an opportunity for improving the entire drug
development process and integrating it as
early as possible with research efforts.
More new drug candidates flowing
through the pipeline will require a new
approach toward product development and
formulation (4). The particular challenge
addressed here is the increasing expectations
placed on development teams to produce
more stable formulations of more
biopharmaceuticals in a shorter period of
time with less material and a finite number
of trained personnel.
Innovative strategies are required to meet
such daunting challenges. The objective is to
find new ways to obtain more information
with less material more quickly than ever
before. Whether that challenge is considered
the last stage of discovery screening or the
first step in product development, the goal is
the same: Identify the critical parameters
that enable the manufacture of a consistent
drug product with suitable stability and
marketability.
The challenge. Many lead molecules fail in
the development pipeline due to inadequate
efficacy models, poor pharmacokinetics,
metabolic instability, low aqueous solubility,
immunogenicity, or unacceptable toxicity.
Because the industry norm (one success in
10 development projects) can be tolerated no
longer in tightening economic conditions,
such problems lead to project termination,
which is the primary determinant in time
and cost overruns. However, this belt
tightening offers an opportunity to improve
the selection criteria at a project’s front-end
rather than continually attempting to rescue
failing projects.
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FEBRUARY 2002
A comprehensive assessment of a drug
candidate’s “developability” would include
its cost-of-goods, process yield, formulation
challenges, and other chemistry,
manufacturing, and control (CMC) issues
critical to bringing a drug to market.
Figure 1 shows that CMC issues are often a
major reason for failure to receive regulatory
approval (3). CMC failures that can derail
further development include inconsistencies
in manufacturing, scale-up difficulties from
bench to manufacturing, variations in
composition, inconsistent purity levels, and
product instability.
The solution. We believe that the drug
development process can be — indeed must
be — more efficient and systematic. By
addressing the endemic inefficiencies in
current systems, drug development can be
accelerated. The “Drug Development
Challenges” sidebar lists seven areas in
which improvement is needed. Such
improvements will increase development
success rates and make the process less
costly and more efficient and timely.
Whereas implementing changes in any of
those seven areas would improve
developmental efficiency, addressing of all
them would be ideal.
Advances in Data Management
Using more sophisticated analytical
methods, modern biopharmaceutical
companies are proficient at data generation.
However, information is often catalogued
and set aside as quickly as it is generated. As
a result, its intrinsic value is frequently lost.
Unless a company makes a concerted effort
to retrieve and analyze that data, it places
itself at a competitive disadvantage.
Poor information management is usually
found in three areas. First, project managers
and senior executives lack the information
necessary to make critical business
decisions. Second, senior technical
personnel who leave the company take with
them needed expertise because no
comprehensive record has been kept of their
knowledge and experience. Third, the
expansive database is unused when it could
be structured to provide a training tool for
new employees. Without coherent and
complete information recovery from data
stockpiles, training is insufficient, reducing
productivity from new personnel.
Information recovery. Despite the perception
that information recovery from stockpiled
Toxicity
22%
Poor CMC
41%
Efficacy
31%
Marketability
6%
Figure 1. Reasons for drug product failure
data is simply a database or software
problem, the issue is more complex.
Establishing a mechanism to track data is
important, but such efforts rarely convert
data into useful information. Only someone
experienced in drug development can
analyze the data sets and construct valuable
summaries. Therefore, we envision a
growing need for development-savvy data
analysis experts who will work on-site to
recover information from company files as
projects move forward.
Although such information retrieval is a
daunting challenge, it will build more
understanding of the corporate knowledge
base and better equip managers with
information on specific projects. Data
analysis experts can partner with educational
professionals to prepare teaching materials
for new employees. Such a data analyst
Drug Development Challenges
For a more efficient system, consider
these changes to your drug
development process:
Improve information management.
Optimize analytical methods.
Borrow from emerging drug discovery
technologies to evaluate product
structure and stability.
Devise effective formulation and stability
design strategies.
Develop improvements and new
technologies in bioprocessing.
Employ new statistical and mathematical
tools.
Rely more on highly automated robotics
systems.
Process Development
Table 1. Medicines in development, either
human clinical trials or waiting for
regulatory approval (2)
Medicines
in Development
Indication
Cancer
Special needs
Heart disease and stroke
AIDS
Mental illness
Alzheimer’s disease
Diabetes
Arthritis
Parkinson’s disease
Osteoporosis
400
200
100
100
100
26
25
19
16
14
would be able to reconstruct much of the
knowledge that leaves with departing
technical personnel. Ultimately, construction
of a new information base (one that contains
data, analyses, and development-directed
reports — not just raw data) will propel
companies that make the effort ahead of
their competitors. Access to that information
by all technical personnel empowers them to
proceed with the commercialization of new
products at a faster pace and with more
confidence than ever before. In addition, a
catalog of such technical reports can
facilitate the regulatory submission process
in a global marketplace.
Integrated Analytical Methods
Another challenge facing development
teams is the lack of capacity for highthroughput quantitative analysis. Obtaining
timely results from analytical groups is
usually a rate-limiting step for process
science units (such as fermentation,
purification, and formulation), as well as
those in manufacturing units.
Despite the analytical armies often
employed in pharmaceutical companies, the
analytical groups are typically stuck doing
things the “old fashioned” way, using
techniques that have been used for decades.
Clearly, with the onslaught of development
projects and the need to make development
faster, new paradigms in the analytical arena
need to be devised and implemented. The
high-throughput technology platforms used
in proteomics can be adapted to drug
development tasks that require quantitative
information.
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BioPharm
FEBRUARY 2002
To devise appropriate characterization
strategies for a new biopharmaceutical drug
candidate, information must be gathered
about the biochemical and physiochemical
properties of the new entity. That
information is also necessary for optimizing
developmental processes such as
fermentation, purification, formulation, and
eventually GMP manufacture. Obtaining the
pertinent information required for rapid and
organized manufacture of a drug product
shortens the development timeline.
Moreover, downstream development
problems would probably be diminished.
Often, no organized strategy for
development activities exists, even for welldefined needs such as the preformulation
studies for biopharmaceutical drug
candidates. In addition, the dramatic
increase in drug candidates will limit
available material more than ever. New
analytical approaches need to consider that
limitation. It may be that the plethora of
analytical methods for characterizing
proteins obfuscates the proper choice of
method and sample conditions (5).
Analytical methods should provide as
much information as possible. For example,
orthogonal techniques can be coupled to the
analytical train to generate additional data
from a given sample. Therefore, the
construction of the development laboratories
of the future must be built around
technologies and methods that provide
detailed, timely information on samples.
Borrowing high-throughput and high-sensitivity
methods. Extended train approaches do not
necessarily address the decreasing
availability of bulk drug material. Therefore,
integrated equipment platforms or suites of
methods linked by an information system
can make use of existing analytical
technology.
The revolution in drug discovery has
produced corresponding innovations in
technology. For example, the conversion of
traditional cell biology methods to
microplate systems has increased
automation and throughput, while requiring
less material per sample. Furthermore, chip
technologies, using structure-recognition
(such as DNA or protein chips) require very
small amounts of material. Finally,
improvements in mass spectrometry (MS)
and its ability to interface with a variety of
separation or high-throughput platforms
(such as capillary electrophoresis, liquid
chromatography, microplates, and
microfluidic devices, for example) provides
the high sensitivity and specificity needed
for preformulation work (6–9).
Integrating high-throughput technologies
into development efforts would increase a
company’s ability to work with small
samples and shorten assay times (per
sample). Consider a few of the technological
innovations that have occurred recently and
determine whether they might be suitable for
your drug development groups, recognizing
the premium placed on accurate and precise
quantitative measurements. Some examples
of methods that might end up in the
development laboratories of the future
include those listed in the “Labs of the
Future” sidebar. Capitalizing on robotics,
miniaturization, and microfluidics
technologies may expedite the development
process using limited quantities of material.
Transfer of any of those technologies to the
development arm of a company could result
in increased throughput, provided the
company is able to generate data that is
suitable for regulatory submissions.
An Efficient Formulation
Even with improved biophysical and
biochemical methods for characterizing the
structure, stability, and function of proteins,
product development for biopharmaceuticals
requires a comprehensive strategy for
moving the product forward to the clinic
with a formulation that is stable and can be
manufactured as easily and inexpensively as
possible. Examining the comprehensive
approach to formulation development taken
by companies can be useful in creating your
own strategy. Although some companies
have detailed, well articulated development
plans, others do not. Until a company
develops such protocols for all process
sciences (not just formulation), its
development process will be inefficient.
Formulation strategies for some companies
employ marginal frozen solutions to initiate
clinical trials, opting to modify the
formulation later if the performance in
humans warrants further development.
Although that strategy may speed the time to
human testing, it provides inadequate data
for describing the performance of the
material during freezing, storing,
transporting, and thawing. A company runs
certain risks in the absence of that data set.
Failure in the clinic could result from poor
Process Development
stability or improper handling rather than an
intrinsic lack of efficacy. In addition, the
strategy may (and almost always does)
require substantial effort in constructing and
evaluating the new formulation, which will
delay future clinical studies.
Other companies have chosen to use
standard or generic formulations, those with
which they have some experience and that
they believe will work for most protein
candidates. Often, such generic formulations
are lyophilized, and the lyophilization cycle
has been well characterized. Given the
current knowledge on protein stabilization
during freeze–drying, a few formulations
should work for a variety of proteins, over a
range of concentrations (11).
Isothermal tests. Most formulation
studies use formal, real-time storage stability
protocols that are acceptable to regulatory
agencies. Given the aggressive timelines of
a high-throughput development (HTD)
strategy, formulation studies are usually
extrapolated from accelerated storage
studies. Accelerated storage studies typically
use isothermal studies. In other words, the
DESPITE expansions
under way at several
pharmaceutical
companies and
contract
manufacturing
facilities, the industry
faces as much as a
fourfold shortage in
capacity.
protein is stored at a fixed temperature.
Depending on the nature of the drug
candidate and its degradation mechanisms,
temperatures up to 50 °C can be used. At
higher temperatures, however, the ratedetermining pathway might be different
from that of the projected storage
conditions.
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BioPharm FEBRUARY 2002
Labs of the Future
Several new technologies may become
standard equipment in the analytical
laboratories of the future.
Chip-based technologies may be used for
protein analysis. For example, Agilent
Protein Chips can reduce electrophoresis
analysis time from days to hours or
minutes using minimal sample volumes
(10).
Phase fluorescence detection
(fluorescence lifetime measurements)
may yield absolute quantities
independent of the measurement
platform or immunity to photobleaching,
turbidity, and other variables that affect
fluorescence intensity measurements.
Disposable test cartridges may perform
multiple tests on one sample, similar to
Nonisothermal stress tests. An alternative
to isothermal testing, investigated in a few
cases, is the use of nonisothermal stress
testing. In this procedure, the sample is
exposed to a linear (or approximately linear)
increase in temperature over time. Samples
are taken at set intervals and assayed for
degradation. Using various algorithms,
Arrhenius parameters can be determined in a
single experiment, whereas at least three
separate isothermal studies would be
required (12,13). This approach has been
used for a pentapeptide and may be
applicable to proteins in the future (14).
Requiring significantly less material, time,
and methodology, nonisothermal analyses
are an attractive method for estimating shelf
life for biopharmaceutical formulations.
By employing both novel stability
protocols and generic formulations for
proteins with similar degradation
mechanisms, the time and material for
formulation development activities can be
reduced. In addition, with use of highthroughput analytical techniques and
efficient information management systems,
extensive information about the drug
product is available more rapidly.
New Bioprocessing Technologies
The shortage in biopharmaceutical
manufacturing capacity in the United States
and Europe is becoming critical. With the
recent success of a number of
biotechnology-derived products, and as
those being implemented in hospital
acute care settings.
Mass spectrometry techniques may be
coupled with multidimensional
chromatography to reduce analysis time
from one day to 1.5 hours.
Low sample volume spectrophotometers
are now available that use as little as
1 L of sample and will be used for
thermal unfolding curves on micrograms
of protein.
Robotic systems for biophysical analysis,
such as with differential scanning
calorimetry may find their place in future
laboratories. Those currently in use allow
up to 100 samples to be analyzed
unattended.
more drugs gain approval, the
manufacturing systems currently employed
will be inadequate to meet the growing
demand for licensed and development-stage
facilities. The 2002 annual material
requirements for the currently marketed
antibody-based products alone is
approximately 550 kg, according to Bryan
Lawlis, chairman of Diosynth RTP,
speaking at the BIO 2001 conference (15).
Similarly, lyophilization capacity may be
insufficient for the large number of
biopharmaceuticals in development as well.
Therefore, the prediction is that, despite
expansions under way at several
pharmaceutical companies and contract
manufacturing facilities, the industry faces
as much as a fourfold shortage in capacity.
In the past three years, close to 75 new
biotech drugs, vaccines, and new indications
have been approved (2). To commercialize
even a fraction of the biopharmaceuticals
currently in development, new paradigms in
bioprocessing technology will be needed.
Many companies will be trying to find
alternatives to freeze–drying
(lyophilization). Methods that allow the
intermediate storage of large amounts of
sensitive biomolecules will be sought.
Fermentation and recovery streams will
need to be more efficient, either through
molecular biology manipulation or by
increased product yield. For example, high
pressure has recently been used to refold
Process Development
proteins from inclusion bodies or
aggregates, which allows damaged material
that used to be eliminated to be processed
further (16). Manufacturing plants are now
being designed for multiple simultaneous
use.
(unpublished work). We envision a growing
role for PLS in optimizing manufacturing
and formulation in the future. Already, it is
being used to understand the
structure–activity relationships important to
governing passive membrane transport (17).
New Statistical Tools
Comprehensive Strategies
Approaches similar to those discussed above
for formulation development can be applied
to other product development and process
science tasks. Purification schemes that
worked for another protein should be tried
for a similar new candidate. Fermentation
work can begin with cell lines and reactor
conditions that worked in previous studies.
DOE software. More companies are using
design of experimental (DOE) software and
statistical packages because they provide
more information from a limited number of
experiments and show subtle interactions
between variables. DOE software also
provides a rationale for the development of a
given formulation.
However, you must be exceedingly
careful with DOE software. Commercial
software packages are not panaceas.
Scientists should still rely on prior
knowledge about the compound of interest,
be aware of the current best approaches in
their field, and understand the complex and
sometimes fragile nature of
biopharmaceuticals. Otherwise, ignorant
choices based on appealing graphs and
tabular outputs will lead to poor or
inappropriate formulation schemes. It is still
our expectation, however, that DOE
packages will be increasingly used within
development groups. If correctly used, such
programs will be a valuable asset in the
overall development process.
Statistical methods. Another computational
approach that could revolutionize drug
development is the use of multivariate
statistics to identify the critical parameters in
a process — even if the data sets are
incomplete or sparse. We have used
multivariate statistical approaches, such as
projection to latent structures (PLS), to
determine the parameters critical to longterm stability during manufacturing. We
have also used PLS to evaluate which large
protein database sequences would lead to
amyloidgenic disease and to identify the
properties that most predict long-term
stability in a series of formulations
Pharmaceutical companies have devoted
enormous resources to speeding drug
discovery. The result has been incredible
innovation leading to combinatorial
libraries, high-throughput screening, and the
new disciplines of proteomics and genomics.
However, similar effort has not been made
in drug development, and that has resulted in
a surge in potential new drug candidates but
limited capacity to move them forward
toward commercialization. The bottleneck is
no longer in research but in product
development. It is our opinion that only a
comprehensive HTD effort can prevent
many valuable lead compounds from sitting
idly on the shelf.
Opportunities are already available for
increased drug development efficiency, such
as high-throughput processing of samples
with minimal material requirements. The
challenge will be to make those analytical
systems more quantitative rather than
qualitative. Many information technology
companies are devoting resources toward
advancing information management
software systems for pharmaceutical
companies. As biopharmaceutical
companies become more sophisticated,
some of the lessons from other hightechnology industries (such as the
semiconductor and computer fields) may be
applicable in the pharmaceutical sector. The
revolution will only come, however, when
management realizes that a crisis is upon us
and that the development schemes of the
past three decades will no longer suffice.
With that realization, high-throughput
development could become a reality.
(5)
(6)
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BioPharm
FEBRUARY 2002
References
(1) A. Dove, “Proteomics: Translating Genomics
into Products?” Nature Biotechnol. 17,
233–236 (1999).
(2) PhRMA Annual Report (Pharmaceutical
Research and Manufacturers of America,
Washington, DC, July 2001).
(3) Pharma 2005: An Industrial Revolution in
R&D (PriceWaterhouseCoopers, New York,
1998).
(4) R. Nayar and M.C. Manning, “HighThroughput Formulation: Strategies for Rapid
(7)
(8)
(9)
(10)
(11)
(12)
(13)
(14)
(15)
(16)
(17)
Development of Stable Protein Products,”
Rational Design of Stable Protein
Formulations, J.F. Carpenter and M.S.
Manning, Eds. (Kluwer Press, New York,
2001).
A. Pandey and M. Mann, “Proteomics to Study
Genes and Genomes,” Nature 405, 996–997
(2000).
B. Zhang, F. Foret, and B.L. Karger, “HighThroughput Microfabricated CE/ESI-MS:
Automated Sampling from a Microwell Plate,”
Anal. Chem. 73, 2675–2681 (2001).
D.I. Papac and Z. Shahrokh, “Mass
Spectrometry Innovations in Drug Discovery
and Development,” Pharm. Res. 18, 131–145
(2001).
A.P. Jonsson, “Mass Spectrometry for Protein
and Peptide Characterisation,” Cell Mol. Life
Sci. 58, 868–884 (2001).
L.F. Kricka, “Microchips, Microarrays,
Biochips, and Nanochips: Personal
Laboratories for the 21st Century.” Clin. Chim.
Acta 307, 219–223 (2001).
L. Bousse et al., “Protein Sizing on a
Microchip,” Anal. Chem. 73, 1207–1212
(2001).
J.F. Carpenter et al., “Rational Design of
Stable Lyophilized Protein Formulations:
Some Practical Advice,” Pharm. Res. 14,
969–975 (1997).
X. Zhan et al., “Exponential Heating in Drug
Stability Experiment and Statistical Evaluation
of Nonisothermal and Isothermal Prediction,”
J. Pharm. Sci. 86, 709–715 (1997).
X. Zhan, G. Yin, and B. Ma, “Determination
of Rate Order for Degradation of Drugs with
Nonisothermal Stability Experiment,” J.
Pharm. Sci. 86, 1099–1104 (1997).
M.L. Lee and S. Stavchansky, “Isothermal and
Nonisothermal Decomposition of
Thymopentin and Its Analogs in Aqueous
Solution,” Pharm. Res. 15, 1702–1707 (1998).
B. Lawlis, paper presented at the BIO 2001
International Convention and Exibition, San
Diego, 23–27 June 2001.
R.J. St. John, J.F. Carpenter, and T.W.
Randolph, “High Pressure Fosters Protein
Refolding from Aggregates at High
Concentrations,” Proc. Natl. Acad. Sci. USA
96, 13029–13033 (1999).
U. Norinder and T. Osterberg, “Theoretical
Calculation and Prediction of Drug Transport
Processes Using Simple Parameters and Partial
Least Squares Projections to Latent Structures
(PLS) Statistics: The Use of Electrotopological
State Indices.” J. Pharm. Sci. 90, 1076–1085
(2001). BP