bio far ma cos

8/4/2019 Bio Far Ma Cos

1/77

Opportunities and Challenges

in modern Biotechnology

by

Prof. Dr. Rainer Fischer


2/77

Kondratieff-Cycles: Key Innovations.

... initiate new industrial and social stages of development

linked world

1900 1950 2000

Steel,Railway,Transport

Internet,

Mobile

Communic.Cycles

EarlyIndustrialisation

1850 1900 1950 2000

Automobil,

Petrolchemistry

Microchip

Automation

Life-sciences

Solar technology

Steem-maschine,Clothing-industry

Innovation

LateIndustrialisation

Service-society

Knowledge-society

HealthAge

E-Technics,

Chemistry

Source: similar in Nefiodowin Capital 1/2 2000


3/77

What is Biotechnology ?

bios = life

teuchos = tool

logos = study of or essence of

e.g. the study of tools from living things/organisms

classical definition Biotechnology is a set of tools that

utilize living things (and more recently, derivatives of living

things) to solve problems or to provide products


4/77


biotechnology is the application of various sciences (i.e.,

immunology, molecular biology, biochemistry, botany, animal

science, etc.) to develop products or to solve problems.

the office of Technology Assessment of the U.S. Congress defines

biotechnology as "any technique that uses living organisms or

their products to make or modify a product, to improve plants or

animals, or to develop microorganisms for specific uses."

the use of living things or parts of living things to create or modifydrugs and other substances

to modify food crops and other macroscopic organisms

to adapt microorganisms to agricultural, medical, or other purposes


5/77


all lines of work by which products are produced

from raw materials with the aid of living things

Karl Ereky 1917

rawmaterials livingsystem product


6/77


recombinant genetic engineering

.using biological process to develop products

G. Steven Burrill 1997

genetic

engineering

raw

materials

living

systemproduct


7/77


protein

bioproduction of drugs so complex

they can only be synthesized in a living system

DNA


8/77



9/77

The Promise of Biotechnology

diagnosing disease

curing disease

nutritious food/feed

healthy food/feed

productive land

feeding the poor sustainable agriculture

healthy environment


10/77

The Breakthrough Experiments in Genetics

Hershey and Chase 1952

T2 bacteriophage: 32P DNA not 35S protein encodes genetic inform.

Watson, Crick, Franklin and Wilkins (1953)

X-ray crystallography

1962 Nobel Prize awarded to three men

Chargaff DNA base ratios

structural model of DNA developed

Messelson and Stahl

14

N/15

N semi-conservative replication confirmed scientific foundation of modern biotechnology based on knowledge

of DNA, its replication, repair and use of enzymes to carry out in vitro splicin

DNA fragments DNA polymerase, DNA ligase, restriction endonucleases.


11/77

Breaking the Genetic Code Finding the Central Dogma

an RNA Club organized by George Gamow (1954)

assembled to determine the role of RNA in protein synthesis

radioactive tagging experiments demonstrate intermediate

between DNA and protein = RNA

RNA moves from nucleus to cytoplasm site of protein synthesis

DNA RNA Protein

transcription translation

genetic code determined for all 20 amino acids by Marshal Nirenberg and

Heinrich Matthaei and Gobind Khorana Nobel Prize 1968

3 base sequence = codon


12/77

Origins of Biotechnology

historical pharmaceutical biotechnology:

Alexander Fleming discovery of penicillin

from bread mold - 1928

Large scale broth tank production of

penicillin for WWII injuries Florey & Chain

modern pharmaceutical biotechnology:

interspecies genetic transplantation

hybridoma tumor cell and leukocyte fusions

heterologous protein production (microbes, animal and plant cells)


13/77

Biotechnology Timelines

1750 B.C. Sumerians use yeast to brew beer

500 B.C. Chinese use mold as an antibiotic to treat boils

1863 Mendel discovers transmission of genetic traits

1906 first early study of genes; term genetics introduced

1919 term biotechnology first used by agriculturalist

1928 Penicillin discovered

1953 Watson and Crick discover

double-helix structure of DNA


14/77

Biotechnology Timelines

1960 first synthetic antibiotic

1965 mouse-human cells successfully fused

1966 genetic code cracked

1973 recombinant DNA technology to cut and paste genes

1975 hybridoma technology (monoclonal antibodies)

1978 insulin gene cloned

1981 first transgenic animal

1983 first transgenic plant (tobacco)


15/77

Biotechnology: last 20 Years

1983 first artificial chromosome

1985 genetically engineered plants field tested

1986 use of microbes to clean up oil spill

1988 first patent for genetically altered animal (transgenic mouse)

1995 first non-viral full gene sequence completed

1997 Dolly the cloned sheep unveiled

2002 mapping of human genome virtually complete

2005 human genome confirmed


16/77

What is Genetic Engineering ?

genetic engineering is the basic tool set of biotechnology

genetic engineering involves:

isolating genes

modifying genes so they function better

preparing genes to be inserted

into a new species

developing transgenes

analysing transgenic organisms


17/77

What is Genetic Engineering ?

Recombinant DNA Technology

Boyer and Cohen, 1973


18/77

Biotechnology Applications

food processing (cheese, beer, dairy products)

production of new and improved crops/foods, industrial chemicals,

pharmaceuticals and livestock

diagnostics for detecting genetic diseases, forensic applications

gene therapy (e.g. ADA, CF)

vaccine development (recombinant vaccines)

environmental restoration & bioremediation

protection of endangered species & conservation biology

plant biotechnology (pathogen and stress resistance)


19/77

Evolution of Biotechnology

Double helix ofDNA structure

DNAcloned

Monoclonalantibodiesproduced

Recombinantinsulin approved

Genetic codeelucidated

A T

C

T

A

T

C

T

G

G

A

G

T

G

T

G

A

A

C

C

PCRreported

Biologicalsapproved for

clinical use

Combinatorialchemistry

Pharmaco-genomics

Gene therapy

Genomics

Humangenomemapped

Human gene cloned

CG GC

T A A T

G C T A

1953 73 75 826165 77 86 1986 - 1999 2000 +


20/77

Waves of Discovery Technologies

1975-1990 The Molecular Stone Age early Genentech, Amgen

1990-2000 Genomics

- Incyte - HGS- Millennium - Celera

1995-2008 Proteomics

- Large-Scale Biology

- Celera

- Oxford Glycosciences

1985-2050 Bioinformatics

2000-2050 Structure-based Design

2002-2040 (High Throughput) Imaging


21/77

Products produced with Biotechnology based enzymes


22/77

The Orgins of Biotechnology

fermentation

tools/sources

grains

yeast

vessels

products


23/77

Industrial Biotechnology

the application of life sciences to conventional manufacturing

and synthesis processes: uses genetically engineered bacteria, yeasts,

plants

usually results in:

lower production costs

more profit - $

less pollution

resource conservation


24/77

Industrial Biotechnology: range of activities

Biobased Products Manufacturing Nanotechnology

Bioenergy and Synthesis Biotech Interface


25/77

Industrial Biotechnology based early stage products

different types of beer & wine

dairy products (joghurt, cheese)

vinegar glycerol

acetone

butanol

lactic & citric acid

antibiotics WWII (Bioreactor developed for large scale production, e.g.penicilin made by fermentation of penicillium)

today many different antibiotics are produced by microorganisms

cephalosporins, bacitracin, neomycin, tetracycline..)


26/77

Fermented Foods and Beverages

long history of fermented foods since people began to settle (9000 BC):

often discovered by accident!

improved flavor and texture deliberate contamination with bacteria or fungi (molds)

examples:

bread

yogurt

sour cream

cheese (chymosin)

wine & beer

sauerkraut


27/77

Beer

barley moistening and

germination; enzymatic

relase of carbohydrates

drying, crushing, and mashing:

further enzymatic release of

maltose, dextrins and proteins

addition of hops, and heat in brew

kettle; clarification

remove hops, add yeast to initiate

alcoholic fermentation

storage (lagering) and packaging


28/77

Wine

grape pressing: must

sterilization & SO2 ; addition of

yeast starter culture

fermentation of must (sugar

content essential for product)

removal of excess yeast

malolactic fermentation

removal of excess yeast

aging & bottling


29/77

Optimization of Yeasts


30/77

Microbial based Products

amino acids to improve food & feed taste, quality or preservation

enzymes (cellulase, collagenase, diastase, glucose isomerase,

invertase, lipase, peroxidase, laccase, pectinase, protease)

vitamins

pigments (melanins)

chemical transformation: substrate + microbial enzyme product

examples: cholesterol steroids (cortisone, estrogen, progesterone) hydroxylation

reaction -OH group added to cholesterol ring


31/77

The Perfect Enzyme (enhanced stability)

sampling nature & screening culture collections

improve available enzyme with rational design

random mutagenesis

natures catalysts are extremely well suited to support life;

they evolved to perform optimally in the context of aliving cell as part of a metabolic network

natural enzymes are usually not so well suited for biotechnology

applications, because of the distinct conditions and different demands

biocatalysis applications depend on methods to tailor nature's

catalysts or redesigning them anew


32/77

The Perfect Enzyme: Exploring Natural Diversity

http://www.pmel.noaa.gov/vents/geology/video.html


33/77

One Example

Diversa: has bioprospecting

agreement with USPS

samples hot pools and geysers in

Yellowstone National Park

finds microbes (extremophiles)

with unique genomes and then uses

gene shuffling to discover newenzymes for industrial applications


34/77

Environmental Biotechnology

using life sciences to clean up pollution

bioremediation using:

microbes

enzymes


35/77

How to make inexpensive sugars in large quantities

development of novel technologies for biobased energy and products

commercial viability

economic and ecological sustainability

sugars are the raw materials or

the crude oils that will

be used in biorefineries


36/77

Renewable Sugar Resources

current:

conventional grain milling operations

near term:

microbial/enzymatic hydrolysis of

cellulosic biomass

(R&D -- enzymes being developed)

medium term:

genetically modified plants to produce more

sugars or starch, larger plants and biomass


37/77

Cellulosic Biomass

plant matter made of tightly bonded sugars and lignin

cellulose is made of sugar building blocks:

but is a tough nut to crack

pre-treatment + enzyme (cellulase) treatment = technological

breakthrough

cellulose after

pre-treatment


38/77

Cellulase: enzymatic conversion of cellulose to sugar

improved cellulase is required to make enzyme conversion of

cellulosic biomass economically viable

DOE/contract with Genencor and Novozymes for R&D

to greatly improve activity of cellulase


39/77

Products that can be made from cellulose & sugars

ethanol (transportation fuel)

polymers PLA, PHA, PDO

fine chemicals

bulk chemicals

commodity chemicals


40/77

The birth of an new Industry

University of California, San Francisco

Genentech Inc. South San Francisco

h i h f h


41/77

The Birth of Genentech

excited by the discovery of Boyer and Cohen,

Swanson contacted Boyer

Boyer agrees to give him ten minutes

meeting lasted 3 hours

1976, Genentech was born

http://www.gene.com/

G h h Pi


42/77

Genentech: the Pioneers

initially faced skepticism from academic and business communities,

however..

1977, produced first human protein (somatostatin) inE. coli

1978, genes for human insulin cloned

1979, human growth hormone cloned

1980, Genentech raised 35 million with IPO

(stock price went from initial $35 to $88 after one hour)

1999- Roche purchases all of Genentechs shares ($2.1 Billion)

2000-Genentech ranks 32 in Fortune Magazines list of 100 best

companies to work for in America

Cl i I li


43/77

Cloning Insulin

Recombinant Pharmace tical Markets


44/77

Recombinant Pharmaceutical Markets

world wide biotech market for recombinant therapeutics:

US$55.5 Mrd in 2007; growth ~40% per annum since 1995

growth will be accelerated by:

genomics, proteomics, bioinformatics

modern platform and enabling technologies:

combinatorial libraries, molecular evolution

improved and safer expression systems

forecast for the market share of

therapeutic antibodies

in 2010 ~US$ 24 billion 59 109 313 7121.336

2.2062.958

3.5674.700

24.000

1995 1996 1997 1998 1999 2000 2001 2002 2003 2010

Recombinant Proteins


45/77

Recombinant Proteins

therapeutics (vaccine, antibodies, cytokines, growth factors,

blood substitutes, enzymes and peptides) for treatment of:

infectious diseases of humans and animals

tumor diseases

autoimmunity

allergies

cardiovascular diseases

inflammation and woundhealing

neurological disorders

diagnostics

enzymes

Types of Biologicals in Development


46/77

Angiogenesis Inhibitors

Antisense

Clotting factors

Soluble receptors

Growth factors

Signaling Proteins

Genetherapy

MoAbs

Vaccines

Other

Recombinant proteinsGrowth hormones

Interferons

Interleukins

Immune-based therapy

9

25

5

59

98

83

12

6

3

4

17

Source: PhRMA 2000. Survey of New Medicines in Development, Biotechnology

Types of Biologicals in Development

5

20

6

4

Drug Discovery Process


47/77

Source: Jones-Grizzle and Draugalis, DICPAnn Pharmacotherapy, 1991.

Drug Discovery Process

DISCOVERY

PLATFORMS

PRODUCT

DEVELOPMENT

Target Identification

Toxicology

In Silico Modeling

Pre-Clinical Development

Compound Screening

Lead Optimization

Human Trials

Target Validation Computational Chemistry

Animal Studies

The Magic Triangle of HTS


48/77

The Magic Triangle of HTS

Costs

reagens

consumables

instrumentation

Speed time/well

wells/day

screens/year

Quality

few false positives

few false negatives

S/N, H/L, Z`-factor

H T S

Making Drugs is hard to do


49/77

Making Drugs is hard to do..

HOW MUCH DOES IT COST?

roughly $ 1000 million

That`s more than it cost to build Queen Mary 2

HOW LONG DOES IT TAKE?

from inception to market is between 10-15 years

It took NASA less time to put a man on the moonTHE ODDS OF SUCCESS?

one in 5,000 developing drugs make it to market

Exactly the likelihood of making a hole-in-one

during any given round of golf.

Fortune Magazine 11-72005 p.77

Time Lines and Costs for Pharmaceuticals Development


50/77

Time Lines and Costs for Pharmaceuticals Development

5000

500

50

5

s

u

b

s

t

a

n

c

es

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 years

drug discovery drug optimization clinical efficicay registration/approv.

50 150 450 500 Mio $

screening

optimisation

toxikology

phase I

phase II

phase III

approval

beeinflubarer Bereich

250 Mio $

Leading Products in the Protein Therapeutics Market (US$mn)


51/77

Leading Products in the Protein Therapeutics Market (US$mn)

1,400MAbAbbott LaboratoriesHumira

1,543InterferonsBiogen IdecAvonex

1,808EPORoche/ChugaiNecoRecormon

2,288CSFAmgenNeulasta

2,455EPOAmgenEpogen

2,506InsulinNovo NordiskNovolin

3,273EPOAmgenAranesp

3,324EPOOrtho BiotechProcrit/Eprex

3,334MAbGenetechRituxan

3,542MAbJahnson&Johnson/

Schering Plough

Remicade70

2005

Sales

Protein

Class

CompanyBrand Name

Global Protein Therapeutic MarketArrowhead Publishers 2007

The greatest Pharmacompanies (2005, in billion Euro)


52/77

The greatest Pharmacompanies (2005, in billion Euro)

0 10 20 30 40

Pfizer, USA

GlaxoSmithKline, GB

Sanofi-Aventis, F

Novartis+Chiron, CH

Johnson & Johnson, USA

AstraZeneca, GB

Merck & Co., USA

Roche, CH

Abbott, USA

Wyeth, USA

Bristol-Myers Squibb, USA

Lilly, USA

Amgen, USA

Bayer+Schering, D

Boehringer Ingelheim, D

Takeda, J

Schering Plough, USA Teva+Ivax, Israel

Daiichi Sankyo, J

Eisai, J

WirtschaftsWoche

Page 51

08/06

And the most dynamic Pharmacompanies (2005, in %)


53/77

And the most dynamic Pharmacompanies (2005, in %)1. Boehringer Ingelheim, D

2. Amgen, USA

3. Teva+Ivax, Israel

4. Novo Nordisk, DK

5. Roche, CH

6. Otsuka, J

7. Merck KGaA, D

8. Novart is+Chiron, CH

9. AstraZeneca, GB

10. Abbott, USA

11. Sanofi-Aventis, F

12. Daiichi Sankyo, J

13. Takeda, J14. Lilly, USA

15. Eisai, J

16. Bayer+Schering, D

17. GlaxoSmithKline, GB

18. Astellas Pharma, J

19. Wyeth, USA20. Schering Plough, USA

21. Johnson & Johnson, USA

22. Merck & Co., USA

23. Bristol-Myers Squibb, USA

24. Pfizer, USA

-10 0 10 20 30

WirtschaftsWoche

Page 51

08/06

Expression Platforms


54/77

MicrobesE.coli & Yeast

Expression Platforms

transgenic animalscell lines

Transgenic plants

Fermentation Production Platforms


55/77

Fermentation Production Platforms

1

Advantages

cGMP, cGLP compliant

biologically contained

upscaling for:

E. coli

yeast animal cells

insect cells

plant cells high cell density

reproducibility

yield optimization

Limitations

initial investment

expert staff required engineering & infrastructure

demands

Fermenatation Facility


56/77

y

Cell Separation and StreamlineTM IMAC


57/77

p

Downstream Processing of Recombinant Proteins


58/77

g

Bacterial Expression Platform


59/77

1

Advantages

rapid cloning and expression

yields: 100-1500 mg/L

low cost

proven technology (FDA)

Limitations

most complex proteins are inactive

no eukaryotic post-translationalmodification

poor post-translational assembly

many proteins are mis-folded, costly

re-folding is required: poor yields

endotoxins

P. pastoris Microbial Expression Platform


60/77

p

1

Advantages

commercial systems

yields: >100mg/L

low cost, defined medium

fermentation ready

proven technology

rapid fermentation

Limitations

intracellular proteins often inactive

tedious clone generation

hyper-glycosylation is possible

MeOH use (toxic, explosive)

Mammalian Cell Expression Platform


61/77

1

Advantages

active complex proteins

more authentic glycosylation

well-defined system

FDA approval

Limitations

expensive infrastructure

high media costs

poor assembly of complex

proteins (sIgA)

long development time

viral contamination

oncogene contamination

Transgenic Animal Expression Platform


62/77

1

Advantages

active complex proteins

accurate glycosylation

yields up to 40g/L milk

herds can be bred

Limitations

very slow (years for cloning)

upscale slow (breeding)

BSE and other pathogens

cloning very labor intensive and yet

to be optimised

host protein contamination

expensive: 100K to 300K$/animal

public ethical concerns

Transgenic Plant Expression Platform


63/77

1

Advantages

low initial investment

medium time scale (months)

unlimited scale-up potential

faster than transgenic animals

very low costs


64/77

Production of recombinant proteins in:

intact plants or Bioreactors

Field Trials with GM Plants


65/77

R & D

product developmentprotein engineering

research &

development

small scale pharmaceutical

production in plant cells

Molecular Farming,upscaled production

large scale production

purification

QC, FDA,

marketing

product/technology

transfer

Traditional Breeding and Genetic Modification (GM)


66/77

humanity has been shaping its environment for millenia

wheat, rice, corn, grape are all the product of breeding

GM permits introduction of desirable traits

all our staple crops are GM through plant breeding

traditional breeding is at its limit

GM gives us new opportunities

Our Food Supply


67/77

world population today is ~ 5 billion

this figure will double by 2030

agricultural land is ~ 1.4 billion hectares

decreasing by erosion, salinization and urban growth

it will be ~ 50% less by 2030

~10 billion people to

feed on half the land

The Worlds most important Crops


68/77

How can we obtain better Crops ?


69/77

1. selection

2. breeding

3. hybridization

4. cloning

5. grafting

6. radiation mutagenesis

7. chemical mutagenesis

8. gene splicing

9. genomics/gene expression

10. tissue culture

Major Inputs in Crop Production


70/77

breeding (hybrids)

mechanization

fertilizers

irrigation

crop protectants

Cro

pproductivity/impro

vement

past present future

information

biotechnology

smart breeding

Plant Biotechnology Platforms and Areas of Interest


71/77

enabling technologies: molecular biology, genetics, biochemistry, cell biology

gene expression, promoters, targeting signals

transformation & regeneration technologies

engineering of input traits:

biotic (pathogen) and abiotic stress (salt, drought, frost, UV, herbicides)

metabolic engineering (carbohydrates, starch, lipids, shapes & colours)

engineering of output traits:

product quality (shelf life, nutraceuticals, functional food)

production of enzymes and speciality chemicals, silk fibre monomers

production of diagnostics, therapeutics, blood substitutes and vaccines

phytoremediation

Engineering Viral Resistance inNicotiana tabacum


72/77

virus infected wild type virus resistant transgenic lineexpressing a molecularpathogenicide

Fungal Resistance


73/77

fungus infected

wild type

fungus resistant

transgenic line

Insect Resistance


74/77

anti-feedant proteins

Bacillus thuringensis d-endotoxins

Bt transformed cotton untransformed cotton

Increase Use of Transgenic Crops


75/77

from one million acres to more than ninety million acres in six years

Soybeans 54%* Cotton 61%*

Corn 25%*

*32 % intended 2002

*74% intended 2002 *71% intended 2002

*= per cent of 2001 actual acres *= percent of intended 2002 acres, USDA

The Future of Biotechnology


76/77

TOOLS

PRODUCTS

TARGETS

Monoclonalantibodies

RecombinantDNA

PCR

Gene therapy

Humanized

monoclonals

Genomics

Combinatorialchemistry

Molecularmodeling

Antisensemolecules

Kinases

Largemolecules

Hormones

Smallmolecules

Ex vivo celltherapy

Liposomaldrugs

Oralavailability

Anemia In vitrodiagnostics

In vivodiagnosticsCancer

CNS disorders AlzheimersHormone

deficiencies Inflammation

VirusesInfections

Targeteddelivery

Blood celldisorders

In vivo genetherapy

Past Present Future

Hot Jobs in Biotechnology


77/77

Rational protein design and in silico structure resolution

High throughput screening (genomics, proteomics, imaging)

Large-scale cell culture

Process engineering and scale-up development

Protein purification and downstream processing

cGMP and validation & regulatory affairs

Bioinformatics

Combinatorial chemistry & biology

Corporate development

bio far ma cos

Documents