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Twenty Thousand Years of Biotech - From “Traditional” to “Modern
Biotechnology”
Chapter · April 2013
DOI: 10.1002/9783527669417.ch1
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Part I
Modern Biopharmaceuticals: Research is the Best Medicine –
Sanitas Summum Bonus
Modern Biopharmaceuticals: Recent Success Stories, First Edition. Edited by Jörg Knäblein.
# 2013 Wiley-VCH Verlag GmbH & Co. KGaA. Published 2013 by Wiley-VCH Verlag GmbH & Co. KGaA.
j3
“In all things of nature there is something of the marvelous.”
Aristotle (384–322 BC)
1
Twenty Thousand Years of Biotech – From “Traditional”
to “Modern Biotechnology”
J€org Kn€ablein
1.1
Biotechnology – The Science Creating Life
“Biotechnology” is a combination of the Greek words bios, techne, and logos, meaning
life, technology, and knowledge. Hence, biotechnology is the technical application of
existing knowledge on bacteria for the weal of humankind. Or, according to a lessimpassioned definition from the University of Hohenheim: “ . . . technical (fermentation) processes applying organisms, cells or parts thereof with the aim to
manufacture products for the pharmaceutical, food or cosmetics industry as well as
sustainable waste reduction in the environmental biotechnology field.”
Even more complex is the 1989 European Federation of Biotechnology (EFB)
definition: “ . . . the integrated application of natural and engineering sciences in
order to technically use organisms, cells, parts thereof and their molecular analogous. Thus, biotechnology deals with the application of biological processes for
technical and industrial production and hence is a very application-oriented science
of microbiology and biochemistry in tight conjunction with technical chemistry and
process engineering.”
As we can see, although definitions of the term “biotechnology” can be somewhat
different, one thing they all have in common: biotechnology improves our daily life
(e.g., biopharmaceuticals or genetically engineered food) – in some cases biotechnology is the only enabler of life!
1.2
The Inauguration of Biotechnology
Since thousands of years people manipulated nature in order to make the
maximum use of it – first biological, then technological, and finally biotechnological
(Figure 1.1). Already eighteen thousand years before Christ, people in the Middle
East had successfully domesticated sheep and deer – later (about 5000 BC) pigs by
the Chinese. At the same time, the Sumerians in Mesopotamia (the area between
Modern Biopharmaceuticals: Recent Success Stories, First Edition. Edited by Jörg Knäblein.
# 2013 Wiley-VCH Verlag GmbH & Co. KGaA. Published 2013 by Wiley-VCH Verlag GmbH & Co. KGaA.
1.3 From “Traditional” to “Modern Biotechnology”
Euphrates and Tigris, in which according to the Holy Bible “milk and honey was
streaming” – today Iraq) were capable of brewing beer as depicted in detail on the
Monument Bleu – a Sumerian stone drawing kept in the Paris Louvre – showing
the process of wheat preparation for beer production. In some years, almost half
of the entire wheat harvest was used for brewing the typical beer (Kasch or Bufa). It
should be mentioned though that these early biotechnologies were very empirical
and the underlying processes far from being reproducible.
Another thousand years later, the Egyptians developed the art of winemaking (Irep)
almost to perfection, and about 3000 BC, the Babylonians were able to brew 20 different
types of beer. In Egypt, in 2600 BC, beer (Henket) was considered a staple food and the
breweries were a royal monopoly – the situation was similar in 2200 BC with rice beer
during the Chinese Hsia Dynasty. Another biotechnology was mentioned around 1700
BC in the “Laws of Sumerian King Hammurabi” (1728–1686 BC; one of the most
influential leaders in the orient): a hand-fertilization technique for date palm trees.
Specific fermentation processes were established in areas in which all required
ingredients were available: wheat beer in Middle Europe (malt, hops, Saccharomyces
cerevisiae), rice wine (rice, Aspergillus oryceae), rice liquor sake in East Asia (rice, koji,
S. cerevisiae), kvass in Russia (wheat malt, rye flour, Lactobacillus spec.), pombe in
South America and Central Africa (millet mash and yeast, Schizosaccharomyces
pombe), and pulque by the Aztecs in Middle America (fruits, Zymomonas mobilis).
People having cows produced yogurt (Streptococcus lactis), kefir (Lactobacillus kefir),
and cheese (e.g., Streptococcus salivarius) – although these products were created
more or less randomly by spontaneous fermentation due to the approximately
500 000 microbes that naturally occur in just 1 ml of milk. In contrast, the
Sumerians developed specific fermentation processes to selectively produce certain
different kinds of cheese. Later (around 250 AD), the “Roquefort” (Penicillium
roquefortii) was shipped to Rome as a “Gallic specialty.”
In contrast, brewing of beer was only developed in the sixth century into a stable and
reproducible fermentation process when monks became more and more interested in
this new type of “liquid bread.” Although during the Lenten season, the monks were
prohibited to eat, according to the sentence “liquida non fragunt ieiunum” (“liquids do
not infringe the abstinence order”), they were allowed to drink – and thanks to that, we
nowadays have very delicious and aromatic strong beers. Today in Germany alone,
100 million hectoliters of beer is brewed annually in compliance with the Bavarian
purity law from 1516 using exclusively barley malt, hops, water, and special yeast
strains. Worldwide, beer is still the top selling biotech product with a yearly consumption of approximately 1.5 billion hectoliters worth more than D 50 billion.
1.3
From “Traditional” to “Modern Biotechnology”
Already at the beginning of the last millennium, “traditional” biotechnologies were
developed in order to produce “high value traits”: in 1276, the first whiskey distillery
was established in Ireland, and two hundred years later certain fermentation
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j 1 Twenty Thousand Years of Biotech – From “Traditional” to “Modern Biotechnology”
processes were optimized using special microorganisms in order to produce
sauerkraut (Leuconostoc mesenteroides) and different yogurt types (e.g., Lactobacillus
bulgaricus). With a size of only a couple of micrometers, the microbes are not visible
to the naked eyes; in 1676, Antonie van Leeuwenhoek (1632–1723) was the first
person to watch a microorganism at a 200 magnification using his self-made
microscope. In 1684, he published his first drawings of the observed microbes [1]
and as a result became scientific member of the very prestigious London Royal
Society – albeit he had never visited a university.
Fantastic discoveries paved the way to modern biotechnology in a fast pace. The
foundation of classical genetics was laid by the English scientist Charles Darwin
(1809–1882) in his revolutionary theories on the principle of natural selection. On
July 1, 1859, Darwin presented his seminal paper on the “Survival of the fittest” [2] on
the development of animals to the Royal Linnean Society, which still prides itself as
the repository of natural history expertise in Britain.
There was very little reaction in the room, but the real furor did not begin until
Darwin published “On the Origin of Species” the following year: “There is a grandeur
in this view of life ... that whilst this planet has gone cycling on according to the fixed
law of gravity, from so simple a beginning endless forms most beautiful and most
wonderful have been, and are being, evolved” are the concluding lines of the
revolutionary book. Darwin, knowing the reaction he was likely to receive, held back
for years from airing and then publishing his theories on the principle of natural
selection – and his fears were correct: he was pilloried by the scientific and religious
establishment of the time. One famous caricature depicted him with the body of a
monkey, so angered were people by the suggestion that they might have descended
from the monkeys rather than having been created in the image of God.
His theories are still rejected by some, notably creationists in the United States,
and are less than welcome in the Middle East. Even in Britain a poll in 2006 showed
that only 48% of the people believed in Darwin’s evolutionary theories.
Today, we honor the man for the 150th anniversary of his presentation to the Royal
Linnean Society, and we celebrate Darwin’s 200th birthday and the 150th anniversary of the publication of “On the Origin of Species.” So, obviously, Darwin is still
causing waves after 150 years, and most likely he would have been happy to get a
fraction of this enthusiasm during his life time. . . .
In 1860, it was the French scientist Louis Pasteur (1822–1895) (founder of
microbiology and biotechnology) who was using for the first time “pure isolates”
(phenotypically identical strains) of Acetobacter to convert alcohol into vinegar. Some
years later, in 1866, it was Johann Gregor Mendel (1822–1884) who showed with his
experiments on hybrid peers that some of the phenotypic characteristics can be
transferred from one generation to the next [3].
1.3.1
Molecular Genetics and Enzymatic Kinetics
The Swiss pathologist Johann Friedrich Miescher (1811–1887) was the first who
stained the “nucleus” of a cell and in 1869 at University T€
ubingen isolated its “nucleic
1.3 From “Traditional” to “Modern Biotechnology”
acid.” Although this was essentially the beginning of molecular genetics only, 40
years later, it was the Danish geneticist Wilhelm Johannsen (1857–1927) who coined
in 1909 the term “genotype” with its inheritable characteristics – the “genes.”
Another important milestone toward modern biotechnology was the kinetic description of fermentation processes, that is, the time it took to convert a certain substrate S
with a specific enzyme E into the product P. Enzymes can catalyze chemical reactions by
factors from 100 million up to 1 trillion (108–1012). Assuming a 1012 enzyme-catalyzed
reaction takes 1 second to be completed, without enzyme it would theoretically take
300 000 years! Obviously, most of our metabolic reactions would not be feasible without
enzymes – hence these biological catalysts enable our life.
The enzymes’ catalytic activity stems from their ability to bring substrates into a
favored steric orientation – the so-called enzyme–substrate (ES) complex, which
reduces the activation energy required to convert the substrate into the respective
product. Since one enzyme can only catalyze the reaction of a restricted number of
substrate molecules (because at one point all active sites are occupied – the so-called
saturation effect), there is a maximum reaction velocity depending on substrate type
and enzyme. Chemical reactions without enzymes obviously do not have such
kinetics as it was shown already in 1913 by the Berlin biochemists Leonor Michaelis
(1875–1949) and Maud L. Menten (1879–1960). From their kinetic experiments,
Michaelis and Menten concluded the existence of an ES complex by measuring
the maximum velocity for enzyme-catalyzed reactions [4]. This was the earliest
evidence (indirectly though) of this phenomenon and therefore today it is called
the “Michaelis–Menten kinetic.”
Applying this knowledge to the fermentation process – by feeding the substrate
continuously (rather than batch) to avoid the saturation effect – it was possible to
control the reaction and maximize the time–space yield in a bioreactor. In the following sections, we will see some examples showing that this was really imperative – in
combination with genetic optimization – to produce the required amounts of vital
and lifesaving substances on a large scale.
1.3.2
Penicillin and Other Lifesaving Antibiotics
The immense importance for humankind of such biotechnological developments is
illustrated nicely in the first example: Alexander Fleming (1881–1955) was working
as a bacteriologist at St. Mary’s Hospital in London when in 1928 he discovered a
clear zone around a fungus, which was (accidentally) growing on an agar dish with
Staphylococcus. Obviously, this fungus produced some agent preventing the bacteria
to grow – and since he named it as Penicillium notatum, Fleming called the
compound Penicillin. When Fleming published his work in the British Journal of
Experimental Pathology [5], he did not realize the huge medical potential of this
compound against a variety of hazardous bacteria: streptococci, staphylococci,
anthrax, diphtheria, glandular fever, and tetanus. Ten years later, in 1938, the
Ukrainian biochemist Ernst Boris Chain (1906–1979) at Oxford University understood the general antibacterial potential of Penicillin.
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j 1 Twenty Thousand Years of Biotech – From “Traditional” to “Modern Biotechnology”
With the beginning of World War II in 1939 all of a sudden there was an enormous
demand for compounds fighting bacterial infections to treat the great number of
casualties. Chain and the Australian pathologist Howard Florey (1898–1998) saw the
great need for antibiotics, the potential of Fleming’s work, and continued where he
stopped. Two years later, Penicillin was used to treat a 43-year-old patient suffering
from a severe staphylococci infection. After a short recovery, the patient died one
month later due to the lack of sufficient amount of Penicillin, although Chain and
Florey tried to recover the substance from his urine – so, obviously there was an
urgent need for improved fermentation processes.
One issue though with Fleming’s strain was that it grew exclusively on surfaces
and yielded only 2 mg Penicillin per milliliter of culture broth. Joining forces with the
National Academy of Science and the fabulous Northern Regional Research Laboratory (NRRL), scientists were now looking for a strain that could also be grown as a
submerse culture. In 1943, Penicillium chrysogenum was isolated with a yield of at
least 40 mg/ml Penicillin. Rather than using a surface fermentation the so-called
deep-tank process was applied and with a developed mutant of P. chrysogenum this
technique yielded 150 mg/ml. The production rate could even be enhanced using
residual water from maize processing as a substrate.
Nevertheless, to further optimize the strains by applying random UV mutagenesis
(evolution in the Petri dish), the War Production Board in the United States assigned
universities and other research laboratories. Eventually, also Stanford University, Cold
Spring Harbor Laboratory, and twenty US companies got involved. Optimized
fermentation processes, mutated strains, and feeding strategies (see Michaelis–
Menten) finally led to a production rate of 1.5 mg/ml Penicillin – from 2 mg/ml in the
beginning that was a factor of 750!
On March 1, 1944, the first large-scale fermentation was run and further
technological improvements led to 90% recovery in a 30 000 l reactor (rather
than 1% in a 1 l flask), increasing the yearly produced doses from 210 million to
6.8 trillion!
Only by the joint research and development (R&D) activities of a large number of
very enthusiastic scientists, it was possible to save the life of up to 1500 people
during that war period. Subsequently, on D-day (June 6, 1944) innumerable victims
were wounded but could fortunately be treated with Penicillin. For this success, and
for their achievements in developing Penicillin, Sir Howard Walter Florey, Boris
Chain, and Sir Alexander Fleming were awarded the Nobel Prize for Physiology or
Medicine in 1945.
To treat a variety of other infectious diseases, a number of additional antibiotics
were developed since then, that is, streptomycin, tetracycline (both from Streptomyces spec.), cephalosporin (Cephalosporium acremonium), and rifamycin (Nocardia
mediterranei). They all have in common their selectivity: these antibiotics specifically
interfere with essential microbial metabolism, but not with any human pathways. As
we will see later, antibiotics are not only lifesaving, they are also essential tools for
molecular biology, genetic engineering, and cloning. Perorating, it should be
mentioned that nowadays optimized fermentation processes, using genetically
1.3 From “Traditional” to “Modern Biotechnology”
optimized high-producing strains, are capable of yielding 20 000 times more
Penicillin in 1 l of culture broth compared to what Fleming’s initial P. notatum did!
1.3.3
The Triumphal Procession of Vitamin C
Another very striking example of biotechnological development is the production of
vitamin C (L-ascorbic acid) at large scale. Since humans and other primates do not
have the essential enzyme L-gulonolactone-oxidase, which is required for biosynthesis of vitamin C, they rely on exogenic supply by food (100 mg/day) to avoid
any deficiency signs.
The most popular vitamin C deficiency is scurvy, leading to general bleeding,
distorted development of connective tissue, gingivitis, and loss of teeth. This phenomenon was known since quite some time for sailors who were not provided with
sufficient vegetables and fruits during their long journeys. This was why around
1900 the German emperor gave the order for all sailors to eat a couple of spoons of
citrate every day, which unfortunately did not have the desired results: rather than
prevention of scurvy the sailors were suffering from diarrhea. Why? Although it is
correct that citrus fruits prevent scurvy, we know today it is their vitamin C content and
not the citrate as was believed at those times. And so, twofold Nobel Prize laureate
Linus Pauling (1901–1994) was eating a couple of grams of vitamin C every day
making use of the protective properties of the antioxidants to catch free radicals
(Pauling is one of only four individuals to have won multiple Nobel Prizes: Nobel Prize
in Chemistry in 1954 and Nobel Peace Prize in 1962. With that he is one of only two
people awarded two Nobel Prizes in different fields – the other being Marie Curie for
Chemistry and Physics – and the only person awarded two unshared prizes). As a
consequence of the high vitamin C diet, Pauling never caught a flew and lived up to
93 years, even though he died of cancer of the urinary bladder as a consequence of
over-acidification of his urine.
In 1933, Tadeus Reichstein (1897–1996) at ETH Zurich was able to chemically
synthesize vitamin C for the first time: in more than ten steps he converted
glucose into L-xylose followed by addition of hydrocyanic acid to form L-ascorbic
acid [6]. Nevertheless, this complex chemical synthesis was obviously not suitable
for large-scale production of vitamin C (although Reichstein received the Nobel
Prize for Medicine in 1950, it was not for the vitamin C synthesis, but rather for
his merits on cortisones). Reichstein and colleagues [7] improved the synthesis by
using sorbose as intermediate. This is gained with 100% yield by the reduction of
glucose with hydrogen at a pressure of 150 atm applying a nickel catalyst [8]. The
next step, which is the chemical oxidation of sorbit into the precursor of vitamin
C, sorbose, is very difficult though, because both asymmetric carbon atoms of
L-ascorbic acid stem from glucose already – hence the key step in vitamin C
synthesis is the regio-specific oxidation. And again, the problem can be solved
using bacteria: as early as 1896, the French chemist Gabriel Bertrand (1867–1962)
had described the microbial conversion of sorbit into sorbose with Acetobacter
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suboxydans (according to the present nomenclature Gluconobacter oxydans). Reichstein used its enzyme sorbitol-dehydrogenase to stereo-selectively convert D-sorbit
into L-sorbose followed by chemical oxidation into 2-keto-L-gulonic acid (2-KLG)
and subsequent water cleavage through acid treatment into L-ascorbic acid,
vitamin C.
Named after these “biotech pioneers,” the method is called “Reichstein-andGr€
ussner” process, which was bought by Roche and marketed the vitamin C compound
Redoxon1 in 1934. Also before, Roche was interested in another method to produce
vitamin C. This procedure to isolate vitamin C from paprika was established by the
Hungarian scientist Albert von Szent-Gy€orgyi Nagyrapolt (1893–1986) who in return
received the Nobel Prize in Physiology or Medicine in 1937.
For years vitamin C was produced according to the process of Reichstein and
Gr€
ussner, until lately by means of genetic engineering a pure biotech process was
developed. Bacteria of the species Erwinia use three enzymes to subsequently
convert D-glucose into 2,5-diketo-D-gluconic acid (2,5-DKG), which in turn
can serve as a substrate for Corynebacterium spec. to produce 2-KLG using its
2,5-DKG-reductase (again, using acid treatment this can easily be transformed
into vitamin C).
Although, on first sight, it seems obvious to carry out a co-fermentation with these
two species, this is not possible for multiple reasons: both strains have very different
demands on optimum pH, temperature, and composition of the media. Hence,
Erwinia (Gram negative) and Corynebacterium spec. (Gram positive) have very
different growth rates and hence cannot be grown in the same bioreactor at the
same conditions; these are just a few of the manifold problems that render a cofermentation impossible. Again, it is biotechnology that can solve the manifold
problems by “metabolic engineering”: the genetic information to express the enzyme
2,5-DKG-reductase is transferred from Corynebacterium into the genome of Erwinia
(Figure 1.2). Applying such genetic engineering procedures, it was possible to
generate a recombinant artificial “Erwinia hybrid-strain” that produces in only 120
hours impressive 120 g 2-KLG per liter fermentation broth with a yield of more than
60%(!), which is then directly converted into vitamin C. In this case of metabolic
engineering, the manufacturing costs could be reduced by a factor of 50 and in the
meantime more than 80,000 ton vitamin C are produced every year worth more than
US$600 million!
From this exciting example, it can be demonstrated how a very complex and
sequential low-yield chemical synthesis can be replaced by a pure “one-pot” highyield biotech fermentation process for cost-effective large-scale production. Using
similar approaches, it was possible at BASF to simplify the eight-step chemical
vitamin B2 synthesis into a one-step fermentation process and at DSM to substitute
a chemical ten-step production of the antibiotic Cephalexin with the one-pot
fermentation. As a result, the production costs could be cut in half and the energy
consumption reduced by even 65%. The same is true for environmental pollution,
because rather than using organic solvents for chemical production, only water is
used for fermentation. Hence, these processes are good examples for “white
biotechnology.”
1.4 A Small Molecule from Bacteria – A Huge Importance for Mankind
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Transformation of glucose into vitamin C is catalyzed by four (subsequent) reactions:
3 enzymes from Erwinia
1 enzyme from Corynebacterium
1
4
2
3
Global Pharma Specialists (GPS)
From: “Modern Biopharmaceuticals ® ,” Knäblein 2005
www.get-gps.net
Figure 1.2 Direct conversion of glucose into vitamin C by artificial bacteria: recombinant hybrid
of Erwinia and Corynebacterium.
1.4
A Small Molecule from Bacteria – A Huge Importance for Mankind
1.4.1
Plasmids: Transformation by Gene Transfer
Now we have seen some striking examples of genetic engineering – but how does
that work? In 1928, the English physician and bacteriologist Frederick Griffith
(1877–1941) performed the experimentum crucis, the key experiment for genetic
engineering, when he worked with two different strains of Streptococcus pneumoniae, the causing agent of pleuropneumonia. One strain, contained in a rough
capsule (R), is harmless to mice, whereas the other, with a smooth capsule (S),
kills them. Injection of heat-inactivated lethal S-strains into mice has no consequence, but in combination with the living harmless R-strains the mixture kills
the animals. The “killing agent” was obviously transferred from the dead R-strain
to the (former harmless) S-strain and transformed it into a lethal strain.
With this experiment Griffith was able to genetically transform one strain into
the other: by transfer of genes from S-strains he created transformed R-strains
with S-genotype and S-phenotype. This was the first case of purposely performing genetic engineering by transferring genetic material from one organism
to another [9].
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Based on this exciting experiment, the Canadian physician Oswald Theodore
Avery (1877–1955) at Rockefeller Institute in New York went one step further: he took
cell extracts from heat-inactivated S-strains, which he purified step by step. All
fractions (cell wall, protein fractions, nucleic acids) were analyzed for their ability to
perform the transformation of R-strains into S-trains. By adding ethanol, a precipitate
was formed, and the transformation did not work anymore. Carbohydrates (e.g., the
capsules) do not precipitate, but nucleic acids do, and protease treatment showed no
effects. Transformation is obviously only possible with intact nucleic acid fractions and
when adding the RNA-degrading enzyme ribonuclease the principle still works.
Finally, Avery performed a chemical test for the DNA sugar deoxyribose, which
resulted in the expected blue color. He then concluded that the “transforming agent”
was not a protein, it was not RNA, and it was not carbohydrate – it was DNA. In 1944,
Avery could identify “DNA” as the carrier for all genetic information when he
transformed bacteria by injecting chemically synthesized DNA to gain new properties and hereditary characteristics [10].
Combining these two smart genetic experiments from Griffith and Avery to
identify DNA as “transforming principle and carrier of genetic information” were the
first molecular genetic experiments in history.
1.4.2
DNA: The Molecule of Life
Another quantum leap in molecular biology was in 1953 when the biochemist
James D. Watson (1928–today) and the physicist Francis H. Crick (1916–2004)
elucidated the structure of “the molecule of life” [11]. As spriritus rectors of the
3D structure of DNA, they herald the start of modern biotechnology: this
molecule was not just compelling in its sheer beauty, but more importantly,
understanding the double-helical structure led to the fundamental mechanisms
of transcription and translation (Figure 1.3). This process of copying DNA,
namely replication, was the “first three-dimensional Xerox machine” (Kenneth E.
Boulding, 1910–1993).
Three years later, Arthur Kornberg (1918–2007; Nobel Prize in Physiology or
Medicine in 1959) isolated the enzyme that synthesizes the molecule of life: DNApolymerase [12]. During that time, scientists were also working on the more
complex structures of proteins: John C. Kendrew (1917–1997) and coworkers
described the structure of myoglobin in 1958 [13] and two years later Max F.
Perutz (1914–2002) and colleagues described the structure of hemoglobin [14].
Both shared the Nobel Prize in Chemistry in 1962 for their “studies on the structures
of globular proteins.”
Also in the field of DNA, one important discovery was followed by the next: the
first plasmid was isolated in 1959 and one year later Franzois Jacob (1920–today;
Nobel Prize in Physiology or Medicine in 1965) and Jacques Monod (1919–1976;
Nobel Prize in Physiology or Medicine in 1965) defined the general architecture of
an operon and mRNA as the carrier/blueprint of entire proteins [15]. Another year
later, Marshall W. Nirenberg (1927–2010; Nobel Prize in Physiology or Medicine in
1.4 A Small Molecule from Bacteria – A Huge Importance for Mankind
j13
By splicing-out the introns, mature mRNA is created to yield in the mature protein of choice
mature protein
nucleus
DNA
nucleotide
growing
polypeptide chain
release of
ribosome
mRNA
RNA
Transcription
by RNA-polymerase
tRNA
intron
Translation
of mRNA into p
protein
exon
subunits of
the ribosome
cytoplasm
t l
Global Pharma Specialists (GPS)
mature mRNA
ribosome starts
reading of mRNA
tRNA loaded with
amino acids
ribosome with growing
polypeptide chain
From: “Modern Biopharmaceuticals ® ,” Knäblein 2005
www.get-gps.net
Figure 1.3 Transcription and translation: in the nucleus DNA is transcribed into mRNA, which is
released into the cytoplasm – mRNA is then translated into protein.
1968) started decoding the genetic alphabet by identifying that the codon UUU on
mRNA level was encoding the amino acid phenylalanine [16]. Now the mystery of
transcription and even of translation was solved, and Watson and Crick, along with
their colleague Maurice Wilkins (1916–2004), were awarded the Nobel Prize in
Physiology or Medicine in 1962.
Then in 1968 “gene scissors,” discovered by Werner Arber (1929–today; Nobel
Prize in Physiology or Medicine in 1978), revolutionized molecular biology, since
these restriction enzymes were capable of specifically cutting bacterial DNA with
specific overhanging nucleotides [17]. These “sticky ends” enabled scientists for the
first time to prepare recombinant DNA. Two years later the “central dogma” of
biochemistry was proven wrong, namely that the genetic flow is unidirectional from
DNA via mRNA to protein: Howard M. Temin (1934–1994; Nobel Prize in
Physiology or Medicine in 1975) and David Baltimore (1938–today; Nobel Prize
in Physiology or Medicine in 1975) discovered the viral enzyme reverse transcriptase
[18] synthesizing cDNA from an mRNA template [19]. This breakthrough discovery
eventually allowed the expression of eucaryotic genes, because the non-translated
segments (introns) in the genome are spliced-out by this process yielding mature,
completely coding templates for protein expression: from reading to writing the
genetic code for mammalian gene expression (Figure 1.4).
In 1971, the Protein Data Bank (PDB) was established at Brookhaven National
Laboratory, New York, and became a repository for protein coordinates, which are
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j 1 Twenty Thousand Years of Biotech – From “Traditional” to “Modern Biotechnology”
The enzyme reverse transcriptase uses mRNA as template to create cDNA. Using restriction
enzymes and ligase, this insert can subsequently be cloned into a DNA expression vector
cleavage
g of insert
with restriction enzymes
mammalian
cell
ligase
recombinant expression
p
vector with
mammalian gene
combination of insert
and plasmid by ligase
mRNA with gene coding
f th
for
the protein
t i off choice
h i
isolation
of mRNA
generation of cDNA by
reverse transcriptase
cleavage of plasmid
with restriction enzymes
introduction of expression
vector into bacterial cell
bacterial cell
Global Pharma Specialists (GPS)
plasmid
plasmid
production
expression of mammalian
gene in bacterial cells
From: “Modern Biopharmaceuticals ® ,” Knäblein 2005
www.get-gps.net
Figure 1.4 Expression of eucaryotic genes: mammalian genes can be introduced into E. coli and
functionally expressed by the bacterial protein synthesis machinery.
shared by scientists worldwide. PDB became a very important tool and the basis for
rational, structure-based drug-design – a prerequisite for the development of modern
(bio)pharmaceuticals [20] (see also Chapters 13, 15–19, 22, 23, 25), [21–49].
In 1973, a new era in biotechnology started with the advent of gene technology,
when Allan Maxam (1942–today) and Walter Gilbert (1932–today) at Harvard [50]
and Frederick Sanger (1918–today; Nobel Prize in Chemistry in 1958 for protein
sequencing and the structural elucidation of insulin [51]) at Cambridge [52]
independently developed a “DNA sequencing method.” For these efforts, Frederick
Sanger and Walter Gilbert (but not his student Allan Maxam) were awarded the
Nobel Prize in Chemistry in 1980.
Combining all these fascinating findings, in 1973, Stanley N. Cohen (1935–today)
and Herbert W. Boyer (1936–today) for the first time recombined in vitro DNA pieces
to a new gene [53].
1.4.3
Immortalized Cells: The Source of Monoclonal Antibodies
At the same time, Georges J.F. K€ohler (1946–1995) and Cesar Milstein (1927–
2002) worked together at the Medical Research Council Laboratory of Molecular
Biology in Cambridge, where in 1975 they discovered a technique to produce
monoclonal antibodies. Previously, to prepare substantial quantities of antibodies,
1.4 A Small Molecule from Bacteria – A Huge Importance for Mankind
scientists had to inject an antigen into an animal, wait for antibodies to form, draw
blood from the animal, and isolate polyclonal (a mixture of different types of)
antibodies. The only way to obtain monoclonal antibodies was to clone lymphocytes, secreting one single form of antibody molecules. Lymphocytes, however, are
short-lived and cannot be cultivated easily. By fusing lymphocytes with myeloma
cells (an accumulation of malfunctioning or “cancerous” cells that grow and
multiply uncontrollably), K€ohler and Milstein obtained hybrid cells synthesizing a
single species of antibody while perpetuating themselves indefinitely [54]. A great
number of modern biopharmaceuticals [20,24] (i.e., therapeutic and diagnostic
proteins) today are antibody-based molecules [22,23,33,34], and this is why the
development of monoclonal antibodies revolutionized medicine and paved the way
for new and target-specific approaches, where pure, uniform, and highly sensitive
protein molecules are used as biopharmaceuticals for diagnosis and therapy. For
their achievements, K€ohler and Milstein were awarded the Nobel Prize in
Physiology or Medicine in 1984.
1.4.4
Insulin: The First Biotech Blockbuster
The recombinant DNA technology of Cohen and Boyer [55,56] enabled them in 1978
to generate the first commercial product: human insulin (Humulin1) expressed in
Escherichia coli. These efforts also led to the first biotech company: on October 15,
1980, “Genentech” went public on New York Wall Street. Fascination about this
modern biopharmaceutical and the huge potential of the new biotechnology made
the stock price jump from US$35–89 in the first twenty minutes: by the evening of
the same day, the market capitalization was US$66 million! [57].
Insulin is a naturally occurring peptide hormone produced by the b cells in the
islets of Langerhans of the pancreas in response to hyperglycemia. Insulin facilitates
entry of glucose into target tissues – such as muscle, adipose tissue, and liver – via
binding to membrane and “facilitated diffusion,” or activation of specific receptors on
these cells (Figure 1.5).
Malfunction leads to diabetes mellitus, a group of metabolic diseases characterized by high blood sugar (glucose) levels, which result from defects in insulin
secretion, or action, or both. In type 1 diabetes, this may be due to b-cell destruction
and in type 2 diabetes to a combination of b-cell failure and resistance of target
tissues to insulin action (insulin resistance). The latter disease can in its early
stages be controlled by low-calorie nonsugar diet and/or treatment with oral
antidiabetic drugs, while in the later stages and type 1 diabetes require insulin
treatment. The World Health Organization (WHO) estimated that some 170
million people suffer from diabetes, a figure that is likely to double by 2030.
Although only a minority of these sufferers actually require daily insulin injections, the current world market for insulin is valued at in excess of US$10 billion.
In this context, for example, for Novo Nordisk (one of the leading insulin
producers), the insulin business is continuously growing and the company
expects an overall growth of about 20% annually.
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facilitated diffusion
porines
p
OO22
Äußere
Membran
inner membrane
respiratory
chain
Atmungskette
f Freiediff
free
i
diffusion
Diffusion
periplasm
Periplasma
Innere
Membran
inner membrane
The easiest kind of passive transport is free diffusion
–
Water, O2, or CO2 can freely pass through the membrane
Facilitated diffusion is enabled by two different membrane proteins
– Transport proteins, for example permeases, facilitate the transport of molecules through
the membrane by conformational changes (e.g., uptake of glucose from blood into blood cells)
– Channel proteins, for example porines,
porines are pores filled with water,
water which mediate
molecules up to 600 Da to pass through the membrane
Global Pharma Specialists (GPS)
From: “Modern Biopharmaceuticals ® ,” Knäblein 2005
www.get-gps.net
Figure 1.5 Membranes of a cell: free and facilitated diffusion. Besides free diffusion of O2, bigger
molecules pass through the membrane by facilitated diffusion via permeases or porines.
In the first 60 years after the discovery of insulin by Frederick G. Banting
(1891–1941; Nobel Prize in Physiology or Medicine in 1923) and Charles H.
Best (1899–1978) in 1921 and the successful treatment of diabetics, only insulin
extracted from bovine or porcine pancreas was available to treat type 1 diabetics [58].
Unfortunately, with the rapid increase in the incidence of diabetes, it was no longer
possible to satisfy the pharmaceutical requirement (estimated to be more than 25
metric tons per year) from animal sources. Furthermore, the animal-extracted
insulin is slightly different from human insulin, which might cause formation
of anti-insulin antibodies and allergic reactions. Porcine insulin, which only deviates
by a single amino acid in position B30 (last amino acid of the B chain; Figure 1.7)
from human, can be converted to “authentically” human insulin in a trans-peptidation reaction, in which the alanine is replaced with a threonine.
The developments in molecular biology and biotechnology opened up for new
possibilities, among these, the biosynthesis of human insulin in E. coli. Insulin is
composed of two disulfide-linked peptide chains referred to as the A-chain and Bchain, and the first recombinant approach used E. coli as host for the expression as
fusion proteins. In a later approach in E. coli, pro-insulin (B-chain-connecting-Cpeptide-A-chain) was expressed, also as a fusion protein. In both of these systems,
the fusion proteins were isolated as inclusion bodies and several chemical steps
were needed for dissolution, cleavage, folding, and formation of disulfide bridges
(Figure 1.6).
1.4 A Small Molecule from Bacteria – A Huge Importance for Mankind
j17
Usually transformation is achieved by introducing foreign plasmid DNA. In nature, these are
vehicles, which are used to easily transfer resistance genes from one bacterium to another
C
Fusion
B
A
DNA coding for pro-insulin
native cell
cell with gene for pro-insulin
plasmid-DNA
C-Peptid
II) di
disruption
ti
III) purification
B-Kette
I) expression
A-Kette
I) CNBr
A-chain
II) sulfitolysis
III) O2; pH10,6
B-chain
I) trypsin
II) carboxy
peptidase B
pro-insulin
i
li
Global Pharma Specialists (GPS)
From: “Modern Biopharmaceuticals ® ,” Knäblein 2005
A-chain
B-chain
ACTIVE INSULIN
(authentical human)
www.get-gps.net
Figure 1.6 Production of recombinant insulin: native cells are transformed with plasmid-DNA
coding for pro-insulin, which is then expressed, isolated, and chemically altered.
Later, a single-chain insulin-precursor with a mini-C-peptide could successfully be
produced (also containing the correct disulfide bridges) and secreted in the yeast
S. cerevisiae. Eventually, other mini-C-peptide insulin precursors of human insulin,
with minimal post-fermentation chemistry and purification, could be achieved with
the S. cerevisiae expression system [31]. The demand for a more optimal treatment of
the patient has called for the design and development of new fast- and slow-acting
insulin analogs and has required alterations of the molecule. For example, by
switching the order of K and P at positions 28 and 29 in the B-chain into P and
K results in a fast-acting Insulin lispro1 (the same is true for mutating P at position
28 into D resulting in Insulin aspart1, whereas fusion of two additional arginines
yields the slow-acting Insulin glargine1 (Figure 1.7)).
Another such example is Levemir1 from Novo Nordisk, an unusual long-acting
insulin product that has just recently gained marketing approval. The major
structural alteration characteristic of this insulin analog is the attachment of a
C14 fatty acid via the side chain of lysine residue number 29 of the insulin B-chain.
This promotes binding of the insulin analog to albumin, both at the site of injection
and in the plasma, which in turn leads to a constant and prolonged release of free
insulin into the blood, resulting in a duration of action of up to 24 hours.
After this short excursion on today’s biotech blockbusters, coming back to 1984,
there was another hallmark discovery: the first 3D structure of a transmembrane
protein, the photosynthetic reaction center from Rhodopseudomonas viridis, was solved.
1.5 Biopharmaceuticals – The Mainstay of Modern Biotechnology
The challenge in solving the structure of this huge (150 kDa) protein was that it
consisted of 11 membrane-spanning, hydrophobic a-helices. Solving the 3D structure was a major breakthrough, since many of the most interesting drug targets are
membrane-bound proteins [59]. Together with his colleagues Johann Deisenhofer
(1943–today) and Hartmut Michel (1948–today), Robert Huber (1937–today), my
PhD supervisor at the Max-Planck-Institute, was awarded the Nobel Prize for
Chemistry in 1988.
1.4.5
Polymerase Chain Reaction: How to Infinitely Amplify DNA
Then there is the advent of a surprisingly simple tool that readily revolutionized
molecular biology and heavily influenced modern biotechnology. In 1983, Kary B.
Mullis (1944–today) invented a process he called polymerase chain reaction (PCR),
which solved a core problem in molecular genetics, namely gene amplification. In
other words, “How to make copies of a strand of DNA of interest?” PCR turns the job
over to the very biomolecules that nature uses for copying DNA as well: two “primers”
that flag the beginning and end of the DNA stretch to be copied and an enzyme,
called polymerase, that walks along the segment of DNA, reading its code and
assembling a copy. To complete the PCR cocktail, a pile of DNA building blocks are
added, which the polymerase needs to make that DNA copy in vitro [60]. This process
revolutionizes biotechnology and nowadays is essential for ample applications in
modern biotechnology, for example, in diagnostic [46,61–65] and drug development
[20,34,48,66–70]. Kary B. Mullis was awarded the Nobel Prize in Chemistry in 1993
for this discovery.
Now we have recaptured some twenty thousand years of biotechnology – from the
very early beginning when people manipulated nature in order to make the
maximum use of it until today. Although the aim is still the same, the methods
have changed dramatically. The main focus of today’s modern biotechnology is
primarily to produce biopharmaceuticals “toward human good,” as stated by Nobel
laureate James D. Watson in one of the author’s books: “The making of pharmaceutical and diagnostic agents in cells has moved from edge to the center of their respective
commercial development. ‘Modern Biopharmaceuticals’ comprehensively describes the
many novel ways cells so are being deployed toward human good” [71]. Some examples of
modern biopharmaceuticals will be discussed next.
1.5
Biopharmaceuticals – The Mainstay of Modern Biotechnology
Biopharmaceuticals are currently the mainstay products of the biotechnology
market and represent the fastest growing and, in many ways, the most exciting
sector within the pharmaceutical industry. The term “biopharmaceutical” was
coined in the 1980s, when a general consensus evolved that it represented a class
of therapeutics produced by means of modern biotechnologies. As we have seen,
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already a quarter of a century ago, Humulin (recombinant human insulin, produced
in E. coli, and developed by Genentech in collaboration with Eli Lilly) was approved
and received marketing authorization in the United States in 1982 [20] followed by
products from Novo Nordisk, sanofi-aventis, and Biocon. As the production of
insulin in bacteria had been already established in 40 000 l scale, the era of largescale mammalian manufacturing still was to come [24].
In fact, mammalian cell culture got popular when highly glycosylated products
like tPA1 (tissue Plaminogen Activator), EPO1 (erythropoietin), and factor VIII
[42] (see also Chapters 19 and 20) became attractive as therapeutics, because
these molecules could not be produced in E. coli. In the 1990s, monoclonal
antibodies as biopharmaceuticals were developed, for example, to treat several
targets for rheumatoid arthritis and cancer. Following the aforementioned
invention of K€ohler and Milstein (which made monoclonal antibodies available
as a pharmaceutical resource and allowed the production of the first monoclonal
antibody Orthoclone OKT31 from Johnson & Johnson in 1986), several other
monoclonal antibody-based biopharmaceutical products have been developed and
marketed.
Although the OKT3 antibody had a murine structure, eight years later the first
chimeric antibody fragment ReoPro1 (Eli Lilly) was approved in 1994 followed by other
chimeric molecules, like Rituxan1/Mabthera1 (Roche/Biogen Idec), Simulect1
(Novartis), and Remicade1 (Johnson & Johnson). In 1997 Roche created Zenapax1,
the first humanized antibody, and successfully launched it in the market. With this
technology trend Synagis1 (Astra Zeneca), Herceptin1 (Roche) [34], Mylotarg1
(Wyeth), Campath1 (Bayer), and many others were developed. Finally, the technology
for fully human antibodies was firstly introduced with Humira1 (Abbott) in 2002 and
meanwhile Vectibix1 (Amgen), Simponi1 (Johnson & Johnson), and Prolia1 (Amgen)
also were on the market.
These biopharmaceutical inventions target certain disease indications, for
example, OKT3, Zenapax, and Simulect were developed for organ rejection prophylaxis, whereas Rituxan was developed for cancer-related treatments and rheumatoid arthritis. The top six products Enbrel1, Rituxan, Remicade, Humira,
Avastin1, and Herceptin alone gained sales of more than US$30 billion in 2009.
Whereas the first four mentioned are biopharmaceuticals to treat rheumatoid
arthritis, the latter products are dedicated for cancer indications. Rituxan, Zevalin1,
and Bexxar1 target non-Hodgkin’s lymphoma, Campath chronic lymphatic leukemia, Herceptin breast cancer, and Avastin, Erbitux1, and Vectibix colorectal cancer.
Currently 31 monoclonal antibodies are approved and marketed for therapeutic
use [22,23,33,34].
To further enhance the bioavailability (especially for tumor penetration), also
smaller human protein mimetics and artificial, non-antibody-binding proteins
based on scaffolds have been invented [23,37,72]. These molecules can be produced
in either bacteria or yeasts as alternative to mammalian cell culture.
Today the market for biopharmaceuticals already represents 10–15% of the total
global pharmaceutical market by value, with a total global sales value of US$86
billion in 2008, and is growing two to three times as fast annually. From 1982 to
1.5 Biopharmaceuticals – The Mainstay of Modern Biotechnology
2010, a total of 131 biopharmaceutical proteins, generated by recombinant DNA
technology, were registered in the US and European markets (not counting the
vaccines, blood products, and other biopharmaceuticals, which are of about the
same number [73]). Looking at antibodies, 31 monoclonals have been developed
and, although the molecular targets still seem to be limited, the biopharmaceutical
market has grown with a compounded annual growth rate (CAGR) of 14%
compared to only 4% for the pharmaceutical market. From 1998 to 2009, the
market size has multiplied by almost a factor of eight from US$14 billion to 105
billion, and it is estimated that the market value will reach US$136 billion in 2015.
Altogether, modern biopharmaceuticals such as Genentech’s human growth factor
Somatropin1 or Amgen’s EPO1 have shown that biopharmaceuticals can benefit a
huge number of patients, and also generate big profits for these companies at the
same time. The single most lucrative product is EPO1, and combined sales of the
recombinant erythropoietin products Procrit1 (Ortho biotech) and Epogen1
(Amgen) have almost surpassed the US$10 billion mark.
Biopharmaceuticals now represent an estimated 30% of the global pharmaceutical industry development pipeline. This growth is driven by considerable
unmet medical need, as these complex products enable binding to specific
targets in ways that are simply not possible with conventional medicines. The
great potential of biopharmaceuticals is represented in the development pipeline:
in 2009, there were 162 monoclonal antibodies, 102 other recombinant proteins,
122 biotechnologically produced vaccines, and 33 other molecules evaluated in
clinical phases I–III.
1.5.1
Modern Biopharmaceuticals in Europe
With a focus on Europe, modern biotechnology and its applications generate almost
2% of the EU’s gross value added, indicating that its importance is comparable to
Europe’s largest industry sector, and the European-dedicated biotechnology industry
directly employs almost 100 000 people, mostly in small- and medium-sized enterprises (SMEs) – however, given biotechnology’s “enabling effect,” employment in
industries using biotechnology products is many times higher. Interestingly, revenues for biotech vaccines jumped from D 65 million in 1996 to D 259 million in 2009,
and for example, in industrial biotech, the EU produces about 75% of the world’s
enzymes, and 20% of the agro industry is related to biotech as well.
Today, the majority of innovative medicine is made available by applying modern
biotechnology to their development and/or manufacturing processes. Beginning
with the technologies used in discovering the cause of a disease, to those used in
diagnostic or therapy development – each and every step of today’s procedures is
based on biotechnology. Biotechnology is no longer limited to the “omics” –
genomics, proteomics, and metabolomics. It also provides and detects the receptors,
antibodies, ligands, antagonists, hormones, and cytokines that keep our bodies in
balance and protect us from harm. Hundreds of millions of patients have benefited
from approved biopharmaceuticals manufactured through biotechnology and gene
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technology to treat or prevent heart attacks, stroke, multiple sclerosis, breast cancer,
cystic fibrosis, leukemia, hepatitis, diabetes, and other (also rare) diseases.
“Modern biotechnology” continues to grow twice as fast annually as the traditional
pharmaceutical industry. Today it is already seven times larger than it was 10 years
ago. Currently, the leading classes of biopharmaceuticals are growth factors for
blood cells. These are used in treating anemia, resulting from a variety of conditions
including chronic kidney disease, chemotherapy, radiation treatments, as well as
from other critical illnesses. For the first time in the history of human healthcare,
biotechnology is enabling the development and manufacture of biopharmaceuticals
for a number of rare and very rare genetic diseases. In total, these illnesses affect
some 20–30 million Europeans and their families. Biotechnology has a major
impact on the provision of safe and effective vaccines against infectious diseases
(see also Chapters 15 and 16) [35,74]. It also provides safer recombinant alternatives
to proteins derived from human blood or tissue.
Section 1.6 is largely based on the transcript of the opening speech at Biotechnica
on October 4, 2010, in Hannover, Germany: “The Economic Impact of the Biotechnology Industry and its Potential to Transform the Pharma Industry” kindly
provided by Peter Heinrich.
1.6
Transformation of the Pharma Industry Through Biotechnology
Biotechnology is setting the course of modern medicine. Biotech innovations help to
reveal disease mechanisms, targets, and drug candidates. In addition, they improve
advanced therapies, clinical trials, diagnosis, personalized medicine, and therapy
monitoring by providing markers, models, platforms, tools, and IT.
It is no secret that big pharma’s R&D engine needs a complete overhaul. Despite a
number of bold efforts to bring big pharma’s R&D back to higher productivity levels,
the pace of innovation remains anemic: the long-term average lags at one new
molecular entity (NME) a year per company. Despite R&D spending at a high of 18%
of revenues, big pharma’s R&D productivity declined by 20% between 2001 and
2007. The cost of bringing a new drug to market currently runs at more than US$2
billion. Personalized medicine is shifting the focus of medicine toward R&D and
biotechnology companies in the interests of patients: before personalized medicine
came in fashion, pharma’s focus along the value chain was in clinical development,
regulatory affairs, reimbursement, and marketing – this is still the case. However,
with the trend for more patient-tailored medicine, the value chain moved back
toward R&D and technology transfer, areas that are the strengths of the biotech
industry. This shift within the value chain will bring the biotech and pharma
industries closer together. And the real question is, whether drug makers can cherrypick enough biotech candidates to fill their new-product baskets or whether big
pharma is just too big to deliver real innovation?
In the 1980s, when big pharma produced blockbusters with much greater
frequency, internal champions often led innovations. These leaders could rally
1.6 Transformation of the Pharma Industry Through Biotechnology
troops across functions and shift the focus of R&D efforts nimbly. Then, the
industry’s quest for repeatability and efficiency, an approach focused on “throughput” and “risk mitigation,” began. Repeatable processes delivered a host of benefits
for big pharma, for example, the industry found a steady source of revenue in
marginally differentiating products and making them “evergreen” through extended
releases or co-formulations. Unfortunately, innovation suffered in the process.
Pharma companies learned that they were in need of a higher degree of medical
differentiation to successfully introduce new products into the market and to meet
the patients’ demand for better and affordable medication. This is not a new idea: in
the 1990s, the pipeline for cancer treatments became crowded with pharma
companies developing chemotherapies, mostly with little therapeutic difference.
The difference came from the biotech industry: instead of becoming “me-too,” the
biotech pioneer company Genentech concentrated on changing the way cancer was
treated. They developed treatments based on humanized monoclonal antibodies – a
technology that most pharma companies considered to be too complicated. Genentech’s researchers focused on understanding tumor biology and set goals to take
patient outcomes to a new level. With its innovative approach, Genentech gained
market leadership.
In the following sections are some facts and indicators, which drive the transformation process of the pharma industry.
1.6.1
The Market as Motivation for Transformation
The top five pharma companies have lost 20% of their market value in the past five
years, while biotechnology companies have gained 18%, as shown for the market
capitalization of the following examples:
Genentech (US$100 million) vs. Pfizer (US$98 million)
Amgen (US$52 million) vs. BristolMeyersSquibb (US$42 million)
Gilead (US$40 million) vs. Eli Lilly (US$37 million).
Comparing the market value of the top five US-pharma companies with the entire
market value of the biotech industry shows that they are almost the same: about
US$290 billion. If the value of those biotech companies is added (which were
acquired by pharma companies), the market value of the biotech industry would be
about US$450 billion and the market capitalization of the top five pharma companies would be down to US$122 billion.
As mentioned before, the biopharma market is growing at 14% CAGR compared
to only 4% of the pharma industry. The global turnover of biopharmaceuticals has
grown between 1998 and 2010 from US$20 billion to US$160 billion (Wood
Mackenzie Product review) [21]. Faced with patent expirations, rising expenses,
competition from generics (see also Chapter 18), and pressure on branded drug
prices, big pharma’s revenue gap could balloon up to almost US$100 billion by 2014.
For the 20 biopharma companies in the world, this represents an annual earnings
decline of 8% (Bain & Company).
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1.6.2
Innovations and Where They do Come From
Of the 25 compounds approved by the Food and Drug Administration (FDA) in
2009 – new biological entity (NBE) and new chemical entity (NCE) approvals without
vaccines – only 8 new drug applications (NDAs) were discovered, developed, and
filed by big pharmaceutical companies (32%). The other 17 compounds (68%) were
subject to alliances, partnerships, and licensing with biotechs, or to acquisition
transactions. Innovations also require huge investments over a long period of
time. According to Roche, to develop one drug, 1 million hours of work needs
to be invested, with about 420 researchers involved, who have to conduct about
6500 experiments. This costs about US$1 billion of investments and takes at least
12 years. In other words, about 5000 human-years are required to get a drug
developed. Looking at the global biotech industry financing, biotech funding increased by 84% to US$62 billion in 2009 from just US$33 billion in 2008.
Interestingly, the biotech funding derived from pharma partnerships unproportionally increased over the last years and accounted for 60% of the biotech funding
in 2009. This funding is expected to further increase and will have a positive impact
on the biotech industry, which will further stimulate the pharma transformation.
1.6.3
Mergers and Acquisitions in the Biopharmaceutical Industry and the Impact
on Innovation
With the innovation burden hanging heavily over the pharma industry, many
pharma companies have started to experiment with new R&D models: GlaxoSmithKline restructured its R&D centers to emulate biotech R&D principles and
hopes to replicate an entrepreneurial culture in a large pharma organization. Eli Lilly
acquired ImClone to in-source innovation from outside the company and then left it
as a standalone unit operating independently, basically as Roche did with Genentech. Pfizer also has set-up a biotech incubator to foster their growth, and meanwhile, AstraZeneca has established a biotech focus, merging purchases Cambridge
Antibody Technology and MedImmune into one “biologics powerhouse.”
The history of the pharmaceutical industry is closely linked to such business
practices of the combination and purchase of corporations. From the creation of the
combined entity of Warner–Lambert in 1955, to the purchase of Warner–Lambert by
Pfizer in 2000, to the recent acquisition of Wyeth, pharmaceutical and biotech
companies have expanded their pipelines, portfolios, and sales forces through
mergers and acquisitions (M&A). M&As have defined the landscape of the pharmaceutical and biotechnology industries, both historically and today. The strategic
targeting behind corporate acquisitions, as well as mergers, is focused on intellectual property (IP), sales force efficiency, streamlining R&D, and reorganizing other
key business areas.
This M&A trend is obviously continuing (including hostile takeovers), because it
seems that big pharmaceutical companies cannot effectively develop their
1.6 Transformation of the Pharma Industry Through Biotechnology
desperately needed new drugs on their own: they rather need to extend their
pipelines with new and highly innovative biopharmaceuticals. Looking for the
ongoing convergence of biotech and big pharma, such mega-acquisitions of mature
biotechnology companies (as also nearly seen with Biogen Idec in 2007) is likely to
continue.
And in fact, in November 2010, sanofi-aventis and Genzyme were still at war:
sanofi-aventis had told Genzyme to stand aside and let the shareholders decide on
whether an acquisition should take place – finally, in February 2011, Genzyme was
purchased for more than $20 billion.
Another current change: Roche’s pipeline review could also set the stage for some
new biotech companies to take shape. According to Bloomberg, the Swiss pharma
giant intends to slim down its pipeline and venture capital (VC) companies are
keenly interested in seeing if some of the most promising programs and scientific
talent could be packaged and launched as independent companies. In fact, Roche
has been assessing VC groups’ interest in financing startups. These new corporate
spinouts are gaining considerable momentum on the global pharma industry.
Earlier GlaxoSmithKline launched Convergence Pharmaceuticals with some
impressive venture-backing and programs that no longer qualified as a core asset.
And more startups like these are expected to take shape in coming months.
It also would not be a first for Roche, which has successfully spun out assets in
collaborative efforts to create Basilea and Actelion. While Bloomberg concentrates
on the potential to expand the Swiss biotech scene with new players, there is no
reason why Roche would not be interested in playing on a global field. The analysts
point to Roche’s cancer and metabolism programs as two areas that would be ripe
for a new company launch.
Some companies like the UK-based Vernalis or big pharma–backed initiatives like
Chorus have established virtual development as a viable and often more effective
and efficient development model. Obviously, there is a clear trend from fully
integrated pharma company (FIPCO) to a virtually integrated pharma company
(VIPCO), outsourcing the different steps of the value chain to third parties, such as
contract manufacturing organizations (CMOs), contract research organizations
(CROs), contract service organizations (CSOs), and biotechs.
Such companies do not just manage costs better by limiting full-time employees,
reducing fixed assets, or clamping down on overheads. Rather, their flexibility and lean
structures help them to quickly move on to the next promising idea – thus driving
innovation. But also the patent situation is accelerating transformation of big pharma:
expiring patents and competition from imitation products are increasing the pressure
on the pharma industry (“bitter medicine for big pharma”), and it is especially
struggling as patents for some best-selling drugs expire and price pressures rise in
the United States and Europe due to clashing of the healthcare systems. A total of 40%
of the patents from the 50 most important pharma products will expire within two
years from 2011 – back in 2007 this was only 15% and in 2002 11%.
For example, the Swiss drugmaker Novartis AG will have to deal with a number of
key drugs such as multi-billion dollar seller Diovan1 losing patent protection over
the next few years. In November 2010, the Sunday newspaper Sonntag said Novartis
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was planning a cost-cutting program similar to cross-town rival Roche Holding AG.
Roche, the world’s biggest maker of cancer drugs, said it would slash 4800 jobs
worldwide, hacking CHF2.4 billion (US$2.5 billion) from annual costs.
One option to compensate for the patent expirations and cope with declining
revenues could be biosimilars and biobetters (see also Chapter 18). Those could also
develop into an option for biotechs; and biosimilars are only at the beginning [21]
(see also Chapter 17).
Interesting to note in this context is a very recent decision of the U.S. District
Court for the Middle District of Louisiana, which is the latest jurisdiction to rule that
brand manufacturers cannot be held responsible for injuries suffered by patients
taking generic versions of their drugs. The court granted summary judgment to
Wyeth and Schwarz Pharma in a case involving their gastroesophageal reflux
disease treatment Reglan1 (metoclopramide HCl). The plaintiff in the case took
a generic version of Reglan from January 1998 to July 2009 and subsequently
developed the neurological disorder tardive dyskinesia, which causes uncontrollable
body movements, particularly around the face. This decision is obviously also true
(and timely) for biosimilars, as within the next five years the first biopharmaceutical
blockbusters, including Aranesp1, Enbrel, and Neulasta1 will go off-patent opening
the market for biopharmaceutical follow-ons.
1.6.4
A Focus on the Opportunities of European Biotech Industry
The current situation for Europeans is like that of their US-based counterparts.
European biotech companies demonstrated considerable resilience in the economic
downturn. The number of public companies decreased by only 4%, from 179
companies in 2008 to 171 in 2009 — a much smaller drop than most industry
watchers had expected. Revenues of publicly traded European companies grew from
D 11.0 billion in 2008 to D 11.9 billion in 2009 — an 8% increase that was well below
the 17% growth seen in 2008.
While several of Europe’s leading companies — including Actelion, Crucell, Elan,
QIAGEN, and Meda — continued to post double-digit revenue growth rates, UKbased Shire saw a significant slowdown on its top line. This was largely the result of
the introduction of generic competitors to its blockbuster drug Adderal1. Excluding
Shire, Europe’s other large companies — those with revenues greater than D 200
million — saw their combined top line expand by a robust 14%. However, smaller
public companies below the D 200 million threshold saw revenues decline by 1%,
dragging down the overall sector’s performance.
As in the United States, R&D expenditures failed to keep pace with revenue
growth in Europe. European public companies’ R&D expenditures were essentially
flat, posting a modest 2% decrease in 2009. This was driven not by a few large
companies, but rather by R&D cutbacks across much of the industry.
Similar to the situation in the United States, close to 60% of public companies
reduced their R&D expenditures in 2009. The cost-cutting helped to boost the
sector’s net income by a remarkable 68%, as combined net loss fell from
1.7 Biopharmaceutical Production – Uncorking Bottlenecks or Wasting Surplus Capacity?
D 913 million in 2008 to only D 88 million in 2009. Innovations come from dynamic
entrepreneurs. In the EU, 23 million SMEs stand for 99% of the European undertakings, 100 million jobs, and 60% of the gross domestic product (GDP).
Of this, D 625 million came from improvement on the bottom line and D 147
million came from the decrease in public company count, since most of the
companies that ceased operations or were acquired during the year were in a
net loss position. Despite slowing revenue growth, Shire was able to deliver strong
growth on the bottom line, and a number of other companies — including Genmab,
Meda, Photomed, Q-Med, and QIAGEN — posted strong increases in net income.
But again according to Bain & Company, for most pharma companies there is no
real rescue in sight. Most companies will find that even shopping for innovation
externally cannot help to close the gap. A recent analysis of 6000 biotech projects
available for late-stage licensing showed that only about 200 are likely candidates for
a large pharma company. Of these, fewer than 100 show potential to become topsellers. Taken together, they account for only about US$30 billion in potential
revenue.
1.7
Biopharmaceutical Production – Uncorking Bottlenecks or Wasting
Surplus Capacity?
The ripe and blooming market of biopharmaceuticals, on the one hand, is exciting,
but, on the other, it became obvious that production capacities for biopharmaceuticals would become a bottleneck and that worldwide fermentation capacities are
limited to produce all biopharmaceuticals needed. In the first years of this millennium, pharmaceutical companies were indeed competing for production slots, as
illustrated in a seminal Nature Biotechnology article in 2002 [75]. Keeping in mind
that the annual demand for a first-generation biopharmaceutical like, for example,
Betaferon is 2 kg vs. 300 kg for, for example, a second-generation antibody like
Genentech’s Rituximab1 (factor of 100!) makes these capacity crunches even more
obvious. As a result, a lot of efforts were spent on alternative expression systems, like
transgenic animals [30] and plants [32,36,76–84]. For example, human anti-thrombin (AT) functions to keep blood from clotting in the veins and arteries of healthy
individuals, but this is required in large amounts as a substitute for patients with
hereditary AT deficiency to prevent life-threatening clots. Human anti-thrombin,
ATryn1, is produced in transgenic goats and harvested from their milk. ATryn was
approved in 2006 by the European Medicines Agency (EMA) for use in preventing
clotting conditions during surgical procedures in patients with hereditary AT
deficiency. In 2009, ATryn received also market authorization by the FDA and
signifies the United States’ first approval for a biopharmaceutical produced in
genetically engineered (GE) animals. But apart from this success, no other biopharmaceutical from either transgenic animals or plants ever made it to the market.
Hence, the biopharmaceutical success story created the need for investments
in much more (traditional) production capacity. The biggest volume drivers for
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mammalian cell culture, in terms of sales and of production amounts, are monoclonal antibodies and derivatives thereof. With the evolution of the product portfolio,
the need for commercial production capacities arose. One example is Amgen, which
bought Enbrel innovator Immunex for US$16 billion. Amgen got the approval for
their Rhode Island facility that was going to produce Enbrel in 2002. The facility
housed eight 8000 l bioreactors, which was not sufficient to provide the market on
the long run. Thus, they used additional capacity from a CMO that helped to serve
the market demand. Having seen such an example of market dynamics, other big
product company organizations (PCOs) decided to timely invest into manufacturing
capacities on their own, or to acquire companies who possessed commercial
manufacturing capacity (n.b., the life cycle of the products runs in parallel to the
life cycle of manufacturing). Many companies have transformed from the stage of
having no good manufacturing practice (GMP) capacity over having own clinical
capacity, to even owners of commercial capacities (see also Chapter 23). PCOs who
heavily invested in mammalian cell culture-manufacturing capacity besides Amgen
were Genzyme Bayer, AstraZeneca with MedImmune, Merck KgaA with Serono,
BMS, Genmab, Pfizer with Wyeth, Eli Lilly with Imclone, Merck Inc. with ScheringPlough, and Roche with Genentech. Roche and Pfizer already faced the situation of
surplus capacities following their respective acquisitions, and as a consequence
Roche closed the brand new 200 000 l facility in Vacaville, CA, USA. Pfizer recently
announced the closure of their new Shanbally plant in Ireland.
Following the life cycle of biopharmaceuticals, there will be a time point for each
product when it will be off-patent in the major markets and the sales and production
volume will decrease (see also Chapter 18). If the innovator company is not able to
compensate the emerging gap, it will have spare capacity up to the dimension by
which the facility gets commercially unprofitable.
Facilities from PCOs lacking a timely pipeline with an adequate facility-fit will be
offered for opportunistic contract manufacturing. Already classical PCOs, like
Abbott, for example, have begun to offer GMP capacity. Most of the existing
large-scale facilities are designed as mono-product plants and that often is an
insurmountable hurdle in transferring approved products from one facility to
another. Nevertheless, some PCO collaborations may occur. Not every company
will find the right partner and in consequence there will be some facilities offered for
sale. Genmab, for example, announced the intention to sell their Brooklyn Park
facility [85].
Awaiting a wave of biosimilars between 2013 and 2018, the PCOs have the chance
to enter this business segment and fill their capacities according to their capabilities
and success, or to outsource the remaining production. CMOs can act as suitable
partners for PCOs as their facilities are designed for multi-product use and they
always try to maximize the capacity utilization. By nature of their business they
possess the right technologies and capabilities.
The currently existing capacity will compete with future trends of higher overall
yields and thus building smaller scale bioreactors or using disposable technologies
according to the fragmentation of certain indications and the titer improvements
that the market has frequently seen with product titers of more than 5 g/l [24,86].
1.8 Conclusion and Outlook
In the future, further improvements in titer and downstream yield will be seen that
lessen the need for building more capacity [87].
Up to 2009, the PCOs held about 1.85 million liters of commercial mammalian
cell culture-manufacturing capacity and therewith 81% of the total capacity of
bioreactors that were equal or bigger than 8000 l. CMOs such as Boehringer
Ingelheim, Lonza, Celltrion, Diosynth, and Sandoz had a total volume of
422 000 l. In 2003, Boehringer Ingelheim invested more than US$330 million
(D 255 million) in their large-scale plant and thereby doubled the Biberach capacity
in Germany, now being the market leader with 180 000 l (since Lonza sold their
80 000 l Singapore facility to Genentech) (see also Chapter 23). Lonza already has
invested in another 80 000 l facility, close to the first one. However, the total CMO
capacity will decline to 380 000 l in 2014 in this forecast as Celltrion decided to use
more than 80% of their capacity for their biosimilars, and Diosynth has been
acquired by the PCO Schering-Plugh (Merck Inc.) keeping their large-scale capacity
in Oss (Netherlands) internally. Also the remaining available capacity from Sandoz
is not clear, as Sandoz seems to predominantly operate in the biosimilars area (see
also Chapter 18). The uptake of Celltrion, Diosynth, and Sandoz into the PCO league
lets the total PCO capacity swell to 2.44 billion liters and leaves the CMO playing
field to Lonza and Boehringer Ingelheim.
Altogether, the FDA approved 44 biopharmaceuticals since 2002, with 32% of
them currently manufactured by CMOs. But also the capacity of the CMOs is not
unlimited, and building stainless steel bioreactors takes 5 years, and investment
volumes of several hundred million US dollars for a green-field plant from
engineering to process qualification [88]. Currently, it looks like that the global
production capacity for biopharmaceuticals is sufficient, but this can change
immediately if new and quantity-demanding treatment paradigms arise on the
horizon.
1.8
Conclusion and Outlook
“Men love to wonder, and that is the seed of science” (Ralph Waldo Emerson, 1803–
1882) – this is basically the driving force and enabler for the continuous advancement of biotechnologies over the last twenty thousand years. People manipulate
nature in order to make the maximum use of it – this started already eighteen
thousand years before Christ, but still holds true for today. The difference though
today is that we need to safeguard to use nature, but not to abuse our planet.
Very early people in the Middle East had successfully domesticated sheep and
deer, and later also pigs by the Chinese. At the same time, the Sumerians in
Mesopotamia were capable of brewing beer, and in some years almost half of the
entire wheat harvest was used for brewing the typical beer (Kasch or Bufa). Egyptians
produced wine (Irep), and the Babylonians were able to brew 20 different types
of beer. Specific fermentation processes were established in areas in which all
required ingredients were available: wheat beer in Middle Europe, rice wine and rice
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liquor sake in East Asia, kvass in Russia, pombe in South America and Central
Africa, and pulque by the Aztecs in Middle America. People having cows produced
yogurt, kefir, and cheese, and the Sumerians even developed specific fermentation
processes to selectively produce certain different kinds of cheese – Roquefort was
shipped to Rome as a “Gallic specialty.” In the sixth century, monks developed beer
brewing into a stable and reproducible fermentation process, which leads to a kind
of “blockbuster of traditional biotechnology,” keeping in mind that worldwide beer is
still the top selling biotech product with a yearly consumption of approximately
1.5 billion hectoliters worth more than D 50 billion.
Later, at the beginning of the last millennium, “traditional” biotechnologies were
developed in order to produce “high value traits” such as whiskey, sauerkraut, and
different types of yogurt; Antonie van Leeuwenhoek makes the respective microbes
visible using his self-made microscope with 200 magnification.
Charles Darwin published his revolutionary theories on the principle of natural
selection, Louis Pasteur converted alcohol into vinegar, Johann Gregor Mendel
showed that phenotypic characteristics can be transferred from one generation to
the next, Johann Friedrich Miescher stained the “nucleus” of cells, and Wilhelm
Johannsen coined the terms “genotype” and “genes.”
Michaelis and Menten postulated the existence of an ES complex and defined the
saturation effect, which subsequently revolutionized fermentation processes to
produce the required amounts of vital and lifesaving substances on a large scale.
For example, the production of Penicillin (initially discovered by Alexander Fleming
with yields of only 2 mg/ml Penicillin of culture broth) can be improved with the help
of Ernst Boris Chain and Howard Florey, based on optimized fermentation
processes with feeding strategies (Michaelis–Menten kinetic), and mutated strains
(evolution in the Petri dish), and finally lead to a production rate of 1.5 mg/ml
Penicillin – from the initial 2 mg/ml this is a factor of 750. On March 1, 1944,
the first large-scale Penicillin fermentation was run and further technological improvements led to 90% recovery in a 30 000 l reactor (rather than 1% in a 1 l flask)
increasing the yearly produced doses from 210 million to 6.8 trillion, which enabled
to save the life of up to 1500 people during the war period, and subsequently, on
D-day innumerable victims who were wounded could fortunately now be treated
with Penicillin. Nowadays optimized fermentation processes, using genetically
optimized high-producing strains, are capable of yielding 20 000 times more
Penicillin in 1 l of culture broth compared to what Fleming’s initial P. notatum did!
Then, another very striking example of biotechnological development is the largescale production of vitamin C (L-ascorbic acid). Reichstein was able to chemically
synthesize vitamin C for the first time, when in more than ten steps he converted
glucose into L-xylose followed by addition of hydrocyanic acid to form L-ascorbic acid.
Later, Reichstein and colleagues improved the synthesis by using sorbose as
intermediate. This was gained with 100% yield by the reduction of glucose with
hydrogen at a pressure of 150 atm applying a nickel catalyst. Using bacteria,
Reichstein used the enzyme sorbitol-dehydrogenase to stereo-selectively convert
D-sorbit into L-sorbose followed by chemical oxidation into 2-KLG, and subsequent
water cleavage through acid treatment into L-ascorbic acid, vitamin C.
1.8 Conclusion and Outlook
Other intriguing experiments follow: combining two smart genetic experiments,
Griffith and Avery identified DNA as “transforming principle and carrier of genetic
information,” Watson and Crick elucidated the structure of “the molecule of life,”
DNA, and Arber isolated restriction enzymes capable of specifically cutting bacterial
DNA with specific overhanging nucleotides. Mullis invented the PCR, which solved
a core problem in molecular genetics, namely gene amplification. With this
biotechnology, it is now possible to indefinitely create copies of a certain DNA
strand, using two “primers” that flag the beginning and end of the DNA stretch of
interest. Temin and Baltimore discovered the viral enzyme reverse transcriptase,
synthesizing cDNA from an mRNA template. This breakthrough discovery eventually allowed the expression of eucaryotic genes, because the non-translated
segments (introns) in the genome are spliced-out by this process yielding mature,
completely coding templates for protein expression. Finally, Cohen and Boyer for the
first time recombined in vitro DNA pieces to a new gene.
Using these new biotechnologies, all of a sudden, it became possible to use
bacteria of the species Erwinia to subsequently convert D-glucose into 2,5-DKG,
which in turn can serve as a substrate for Corynebacterium spec. to produce 2-KLG
using its 2,5-DKG-reductase (acid treatment is used to transform it into vitamin C).
Metabolic engineering yields a recombinant artificial Erwinia hybrid-strain, which
produces in only 120 hours an impressive 120 g 2-KLG per liter fermentation broth
with a yield of more than 60%. The manufacturing costs could be reduced by a factor
of 50 and in the meantime more than 80,000 t vitamin C are produced every year
worth more than US$600 million!
The recombinant DNA technology of Cohen and Boyer enables them also to
generate the first commercial product: human insulin (Humulin) expressed in
E. coli. Although, in the first 60 years after the discovery of insulin by Banting and
Best in 1921, successful treatment of diabetics could only be achieved with insulin
extracted from bovine or porcine pancreases, porcine insulin could now be converted
to “authentically” human insulin in a trans-peptidation reaction. But recombinant
biotechnologies now opened up for new possibilities, among these, the biosynthesis
of human insulin in E. coli: single-chain insulin-precursor with a mini-C-peptide
could successfully be produced (also containing the correct disulfide-bridges) and
secreted in the yeast S. cerevisiae. With minimal post-fermentation chemistry (and
purification), fast-acting Insulin lispro, Insulin aspart, slow-acting Insulin glargine,
and long-acting Levemir could be created.
K€ohler and Milstein discovered a technique to produce monoclonal antibodies, and
in fact, a great number of modern biopharmaceuticals (i.e., therapeutic and
diagnostic proteins) today are antibody-based (and hence target-specific) molecules,
with the first monoclonal antibody OKT3 being marketed in 1986. Although this
antibody had a murine structure, eight years later the first chimeric antibody fragment
ReoPro was approved in 1994 followed by other chimeric molecules, like Rituxan/
Mabthera, Simulect, and Remicade. In 1997, Zenapax was the first humanized
antibody, followed by Synagis, Herceptin, Mylotarg, and Campath. Finally, the
technology for fully human antibodies was firstly introduced with Humira in
2002 and meanwhile Vectibix, Simponi, and Prolia were on the market as well.
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As mentioned before, currently 31 monoclonal antibodies are approved and
marketed for therapeutic use, and the top six products Enbrel, Rituxan, Remicade,
Humira, Avastin, and Herceptin alone gained sales of more than US$30 billion
in 2009.
To further enhance the bioavailability (especially for tumor penetration), also
smaller human protein mimetics and artificial, non-antibody-binding proteins
based on scaffolds, have been invented. The great potential is represented with
an impressive 162 monoclonal antibodies in the development pipeline (in clinical
phases I–III).
Altogether, today the market for biopharmaceuticals already represents up to
15% of the total global pharmaceutical market by value (and an estimated 30% of
its development pipeline) with a total global sales value of US$86 billion in 2008.
From 1982 to 2010, a total of 131 biopharmaceutical proteins (including monoclonal antibodies) were registered in the US and European markets with a CAGR
of 14% compared to only 4% for the pharmaceutical market. From 1998 to 2009,
the market size has multiplied by almost a factor of eight from US$14 billion to
105 billion, and it is estimated that the market value will reach US$136 billion in
2015. Modern biopharmaceuticals such as Somatropin or EPO1 have shown that
biopharmaceuticals can benefit a huge number of patients, and also generate big
profits for these companies at the same time. The single most lucrative product is
EPO1, and combined sales of the recombinant erythropoietin products Procrit
and Epogen1 have almost surpassed the US$10 billion mark.
At the same time though, it has also become obvious that big pharma’s R&D
needs to be transformed to higher productivity levels again, because despite 18%
spending, R&D productivity declined by 20% due to a lack of in-house innovations.
Strikingly, the entire pharma market is only growing at 4% CAGR, whereas
biopharma grows at 14% – hence, now the focus is on in-licensing. But there is
no real rescue and most companies will find that even shopping for innovation
externally cannot help close the gap, since a recent analysis of 6000 biotech projects
available for late-stage licensing showed that only about 200 are likely candidates for
big pharma. Of these, fewer than 100 showed potential to become top-sellers, and
that they account for only about US$30 billion in potential revenue.
A total of 25 compounds were approved in 2009, of those, only 8 were discovered,
developed, and filed by big pharmaceutical companies alone. The rest (17 compounds) came from alliances, partnerships, licensing with biotechs, or acquisition
transactions. Many pharma companies now have new R&D models to emulate
biotech principles, for example, acquiring complete biotechs and leaving them as a
stand-alone unit operating independently. Another obvious trend to participate from
external know-how and to share risks is the transformation from FIPCOs to
VIPCOs. Since outsourcing continues to swell, the different steps of the value
chain are contracted to third parties, such as CMOs, CROs, CSOs, and biotechs.
Also the patent situation is accelerating the desperately needed transformation of
the pharma industry: expiring patents and competition from imitation products are
increasing. Currently, 40% of the patents from the 50 most important pharma
products are expiring. Back in 2007 this was only 15%, and in 2002 it was only 11%.
References
Within the next five years the first biopharmaceutical blockbusters, including
Aranesp, Enbrel, and Neulasta, will go off-patent. One option to compensate for
the patent expirations and the declining revenues could in fact be biosimilars and
biobetters, which again brings big pharma and biotechs closer together. This would
guarantee that, in the future, we will have innovative modern biopharmaceuticals
that in many novel ways are being deployed toward human good.
Having said that, we must support innovative technologies, further expand its
applications, and use the impetus that modern biotechnology gives us, leading to
better lives and a sustainable economy. In the words of Max Planck (1858–1947)
“How far advanced Man’s scientific knowledge may be, when confronted with
Nature’s immeasurable richness and capacity for constant renewal, he will be like a
marveling child and must always be prepared for new surprises,” we will definitely
discover more fascinating biotechnologies. In fact, this should be our focus,
because, then, at the dawn of the new millennium, for the first time we could
yield large-enough amounts of biopharmaceuticals to treat everybody on our planet!
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25 See also Apeler, H. (2005) Producing
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26 See also Weber, W. and Fussenegger, M.
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27 See also Rose, T. (2005) Alternative
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28 See also Yallop, C., Crowley, J., Cote, J.,
Hegmans-Brouwer, K., Lagerwerf, F.,
Gagne, R., Martin, J.C., Oosterhuis, N.,
and Opstelten, D.-J., and Bout, A.
(2005) PER.C61 Cells for the
manufacture of biopharmaceutical
proteins, in Modern Biopharmaceuticals –
Design, Development and Optimization,
vol. 3, part IV (ed. J. Kn€ablein), Wiley-VCH
Verlag GmbH, Weinheim,
pp. 780–803.
29 See also Birch, J.R., Mainwaring, D.O., and
Racher, A.J. (2005) Use of the glutamine
synthetase (GS) expression system for the
rapid development of highly productive
mammalian cell processes, in Modern
Biopharmaceuticals – Design, Development
and Optimization, vol. 3, part IV (ed.
J. Kn€ablein), Wiley-VCH Verlag GmbH,
Weinheim, pp. 809–830.
30 See also Echelard, Y. (2005) The first
biopharmaceutical from transgenic
animals: ATryn1, in Modern
Biopharmaceuticals – Design, Development
and Optimization, vol. 3, part IV (ed.
J. Kn€ablein), Wiley-VCH Verlag GmbH,
Weinheim, pp. 995–1016.
31 See also Andersen, A.S. and Diers, I.
(2005) Advanced expression of
biopharmaceuticals at industrial scale, in
Modern Biopharmaceuticals – Design,
Development and Optimization, vol. 3, part
IV (ed. J. Kn€ablein), Wiley-VCH Verlag
GmbH, Weinheim, pp. 1033–1042.
32 See also Klimyuk, V., Marillonnet, S.,
Kn€ablein, J., McCaman, M., and Gleba,
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33 See also Hempel, J.C. and Hess, P.H.
(2005) Contract manufacturing of
biopharmaceuticals including antibodies
or antibody fragments, in Modern
Biopharmaceuticals – Design, Development
and Optimization, vol. 3, part IV (ed.
J. Kn€ablein), Wiley-VCH Verlag GmbH,
Weinheim, pp. 1083–1142.
34 See also Gutjahr, T. (2005) The
development of Herceptin1: paving the
way for individualized cancer therapy, in
Modern Biopharmaceuticals – Design,
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