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Medical biotechnology
- an overview
Maria Judit Molnar
Institute of Medical Genomics and Rare Diseases
Semmelweis University
Budapest, Hungary
TOPICS
Biotechnology in medicine
Milestones in medical biotechnology
Achievements, paradigm shifts
Current trends and research
Conclusions
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Biotechnology in medicine
Biologic
medicinal
product
=
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Definition of biologics
„A biological medicinal product is a product, the active substance of
which is a biological substance. A biological substance is a
substance that is produced by or extracted from a
biological source and that needs for its characterisation and the
determination of its quality a combination of physico-chemicalbiological testing, together with the production process and its
control.”
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COMMISSION DIRECTIVE 2003/63/EC, 3.2.1.1. b,
Types of biologics
Biologics
Blood or blood components
Recombinant DNA
products
Advanced therapy
medicinal products
Therapeutic proteins
Somatic cell therapy
Monoclonal antibodies
Gene therapy
Fusion proteins
Tissue engineering
Xenotransplantation
Vaccines
Allergenic products
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COMMISSION DIRECTIVE 2003/63/EC, 3.2.1.1. b,
Biologics vs. Small molecule
drugs
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Source: Cecil Nick, TOPRA 2010
Biologics vs. Small molecule
drugs
Small molecule
drugs
Biologics
Extracted from biological source or
recombinant DNA techology
Production
Chemical synthesis
Molecular weight
< 10 kDa
Administration route
Mainly oral route
Mainly parenteral route
Immunogenicity
Not an issue
Could elicit immunological
response
Follow-up molecules
Generics
Biosimilars
> 10 kDa
IN SUMMARY, biologics are protein-like macromolecules.
 Potential risk: immunogenicity
 Parenteral administration
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Biotechnology in medicine
Milestones in medical
biotechnology
Achievements, paradigm shifts
Current trends and research
Conclusions
8
Major milestones in medical
biotechnology
1855
Escherichia coli (E. coli) bacterium discovered (later becomes a major
tool for biotechnology).
1973
Cohen and Boyer develop genetic engineering techniques to "cut
and paste" DNA and reproduce the new DNA in bacteria.
1975
Köhler and Milstein: hybridoma technology (Nobel prize 1984).
1977
Genentech scientists and their collaborators produce the first human
protein (somatostatin) in a bacterium (E. coli).
1982
Eli Lilly and Company markets recombinant human insulin - the first
such product on the market.
1986
First therapeutic monoclonal antibody approved by FDA (muromonab
for organ transplant rejection prevention).
2009
First tissue engineered product approved in the EU (ChondroCelect).
2012
First gene therapy approved in the EU (Glybera).
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Recombinant DNA products
E. coli
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Recombinant DNA products
manufacturing process
Cell lines
•Prokaryotic (e.g. E. coli)
•Eukaryotic (e.g. Chinese Hamster Ovary)
Multi-step manufacturing process
1.
2.
3.
4.
5.
Cell line genetic cloning and selection
Upscale (= fermentation)
Downscale (= purification)
Formulation
Fill and finish
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Biotechnology in medicine
Milestones in medical biotechnology
Achievements, paradigm
shifts
Current trends and research
Conclusions
12
Important therapeutic fields of
recombinant DNA products
Oncology
Psoriasis
Autoimmune diseases
Rheumatology
Inflammatory bowel diseases
Endocrinology
Ophtalmology
Cardiology
Neurology
Transplantation medicine
Infectious diseases
Asthma
Haematology
Intensive care
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ENDOCRINOLOGY
Insulin
From the 1920’s
insulin was derived from porcine / cattle
pancreas for therapeutic use (organotherapy)
1982
first recombinant insulin on the European
market
2014
multiple recombinant insulin products on the
market
Rapid-acting
Short-acting
Intermediate-acting
Long-acting
Pre-mixed
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ENZYMES
Gaucher’s disease and imiglucerase
Gaucher’s disease
• lysosomal disease
• deficit of the enzyme glucocerebrosidase
• accumulation of glucocerebrosides
• autosomal recessive trait
Therapy of Gaucher’s disease
• enzyme substitution therapy
• e.g. imiglucerase
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Pompe Disease
Muscle weakness, atrophy
Alpha glycosidase deficiency
Autosomal recessiv
Source: International Society for Mannosidosis & Related Diseases
Monoclonal antibodies
(abbreviated as mAbs)
„bind to a corresponding antigen in a highly specific manner…”
„…like a key
in a keyhole”
TARGETED
THERAPY
Schneider, Curr Pharm Biotechnol., 2008
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Monoclonal antibodies
mechanism of action
antagonism
apoptosis
CDC
Complement
dependent cytotoxicity
Lutterotti & Martin, Lancet Neurol, 2008
ADCC
transporter
Antigen dependent
cellular cytotoxicity
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ONCOLOGY
Trastuzumab
Conventional chemotherapy
• like „carpet bombing” –> frequent adverse events
Monoclonal antibodies (mAbs)
• targeted therapy
 better outcomes
 less adverse events
An example from oncology: Trastuzumab
• indication: HER2 positive metastatic breast cancer
• HER2 is a type of growth factor receptor
• HER2 positive histology: in approx. ¼ of the cases
• trastuzumab targets HER2 and kills cancer cells
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Source: Herceptin®, European Public Assessment Report
ONCOLOGY + AUTOIMMUNE
DISEASE
CD20
• antigen present on mature B cells
• not present on pre-B cells and plasma cells
• B cells are key players in immune responses more specifically in
humoral immunity
Rituximab
• monoclonal antibody (mAb)
• targeting CD20 with high specificity, and depletes B cells
Indications of rituximab
• Oncological diseases
• non-Hodgkin’s lymphoma
• chronic lymphocytic leukemia
• Autoimmune diseases
• rheumatoid arthritis
• specific types of vasculitis
Source: MabThera®, European Public Assessment Report
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AUTOIMMUNE DISEASE
TNF-alfa
• cytokine
• plays important role in inflammatory processes
Etanercept
• fusion protein
• Fc part of a mAb + 2 pieces of TNF-receptors
• targeting TNF-alfa
Indications of etanercept
• rheumatoid arthritis
• juvenile idiopathic arthritis
• ankylosing spondylitis
• psoriatic arthritis
• plaque psoriasis
• paediatric plaque psoriasis
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Source: Enbrel®, European Public Assessment Report
Vaccines
Production
 derived from natural source
 recombinant DNA technology
Prophylactic vaccines
 against infectious diseases
 e.g. Hepatitis B vaccination
Therapeutic vaccines
 e.g. cancer vaccines
 intensive research is ongoing
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Blood and blood components
Whole blood
Blood components
 red blood cells
 white blood cells
 plasma
 clotting factors
 platelets
For the treatment of
 anaemia
 thrombocytopenia
 clotting deficiencies
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Allergenic products
Application
 used for specific immunotherapy (SIT)
 in vivo diagnosis of immunoglobulin E (IgE)-mediated allergic diseases
E.g. Prick skin allergy test
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Gene therapy
„contain genes that lead to a therapeutic effect. They work by
inserting
'recombinant' genes into cells,
usually to treat a variety of diseases,
including genetic disorders, cancer or long-term diseases. A recombinant gene is
a stretch of DNA that is created in the laboratory, bringing together DNA from
different sources”
• First gene therapy approved in EU
• Recurrent pancreatitis (lipoprotein lipase deficiency)
• Vector: adeno-associated virus
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Source: European Medicines Agency homepage
The history of the gene therapy
1977 - A gene was successfully delivered into mammalian cells
1990 - First human gene therapy was approved: SCID
1999 - J. Gelsinger with OTC deficiency died from organ failure
after gene therapy
2000 - A. Fischer cured children with SCID using retroviral vector,
2 of the children developed leukemia. FDA halted the use
of retroviruses in the US
2006 - Patients was successfully treated with metastatic
melanoma using killer T cells genetically retargeted to
attack the cancer cells
2006 - Succesfull gene-based th. for the treatment of HIV:
lentiviral vector for delivery of an antisense gene againts
HIV envelope
2009 - Researchers succeeded at halting adrenoleukodystrophy,
using a vector derived from HIV to deliver the gene for the
missing enzyme
The number of gene therapeutical clinical trials
Phase I
N:1023
Phase I/I
N:342
Phase II
N:1237
Phase II/III N:214
Phase III
2006 N:1064
N:562
2012 N:3398
General applications of therapeutic gene transfer
1. Molecular therapy for genetic diseases
2. Establishment of a stable gene reservoir as a
source of therapeutic proteins in non - genetic
diseases
3.. Destruction of malignant cells in neoplasias
4. DNA vaccines
Gene delivery
Vectors
Strategies
Viral
Retro
Herpes
Adeno
AAV
Lenti
Nonviral
Plasmid
Arteficial Chr
Liposomes
DNAsomes
Nanoparticlees
Molecular therapies
DNA modulating therapies
 Gene replacement – to replace the defected gene
 Gene transfer – to upregulate therapeutic protein
 Gene editing – to correct the defected gene
 Gene shifting – to upregulate healthy mtDNA molecules, change ratio of
heteroplasmy
RNA modulating therapies
 Exon skipping by antisense oligonucleotides
 Exon inclusion by antisense oligonucletotides
 Mutant RNA removal
 Inactivation of the mutant mRNA by RNAi
 Destruction of the mutant mRNA by ribozymes
Protein modulating therapies
 Neutralization of the mutant protein
 Regulating the level of the haploinsufficient protein
 Upregulating compensatory molecule
The Kaplitt Experiment
Tp “calm down” the overactive neurons of the subthalamic nucleus which
gives rise to signals that cause bradykinesia and tremor in Parkinson’s
disease
EXON SKIPPING
•




A mutation-specific therapy
Providing personalized medicine
Simultaneously may correct all isoforms
Maintains the original tissue-specific gene regulation
The antisense compounds inducing exon skipping are small
synthetic, and highly sequence-specific
Exon skipping in Duchenne MD
13% of DMD patients: correct deletions of 50, 52, 45-50, 48-50, 49-50exons
Progress in AON exon skipping therapy in DMD
Timelines
PRO051
Prosensa/GSK
2OME AON
2007 2008 2009 2010 2011 2012 2013
Ph I
Im.
Ph I/II
Systemic adm, iv, weekly
Ph I/II
Study extension.
Ph I
Non ambulant
Ph II
Dosing
Ph III
Efficacy
Eteplirsen
AVI
PMO AON
Ph I
Im.
Ph I/II
Syst. admin. sc, 3 w
Ph I/II
Dosing
Results: Lancet 2011 Aug 13; 378(9791):595-605
P15
Pre P15
P17
P17
P18
P19
P18
Post
Post Pre
Post
Pre
Post Pre Post Pre
• 7 patients responded to treatment dose higher than 2 mg/kg
• Mean dystrophin fluorescence intensity changed from 8·9% to 16·4%
• The cytotoxic T cells in the post-treatment muscle biopsies decresed
in the two high-dose cohorts
• In future studies higher doses of eteplirsen for longer periods of time
will be administered
Somatic cell therapy
„contain cells or tissues that have been manipulated to change their
biological characteristics. They can be used to cure, diagnose or prevent diseases”
• approved in the EU (2013)
• autologous peripheral blood
mononuclear cells activated
with PAP-GM-CSF
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Sources: European Medicines Agency homepage; Barar & Omidi, BioImpacts, 2012
Stem cells
„cells with self-renewing
capacity i.e. the capability to generate daughter cells
multi-lineage differentiation capacity. Stem cells are capable of
and
proliferation as stem cells in an undifferentiated form”
Embryonic stem cells
Adult or somatic stem cells
• Bone marrow (hematopoietic) stem cells
• Mesenchymal stromal / stem cells
• Tissue-specific progenitor cells with a more restricted differentiation capacity
responsible for normal tissue renewal and turnover (neurons, intestine, skin, lung
and muscle)
Induced pluripotent stem cells
Genetically modified stem cells
Source: EMA/CAT/571134/2009
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Stem cells
Potential applications of stem cells
 metabolic, degenerative and inflammatory diseases
 repair and regeneration of damaged or lost tissues
 treatment of cancer
Umbilical cord stem cell
preservation
e.g. ReNeuron’s ReN001
Phase 1 trial is ongoing, phase 2 trial
application submitted
For the treatment of ischemic stroke
Source: EMA/CAT/571134/2009; ReNeuron homepage
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Tissue engineered products
cells or tissues that have been modified
repair, regenerate or replace tissue”
„contain
so they can be used to
ChondroCelect
o first advanced therapy medicinal product in the EU in 2009
MACI
o matrix autologous chondrocyte implantation
o approved in EU in 2013
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Source: European Medicines Agency homepage
Regenerative medicine
3D printing
Bone grafts
Blood vessels
Ear printing
Printing of a kidney
Liver printing with hepatocytes (awaited for 2014)
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Biotechnology in medicine
Milestones in medical biotechnology
Achievements, paradigm shifts
Current trends and research
Conclusions
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Personalized medicine
"the right patient with the right drug at the
right dose at the right time."
To date more than 100 drugs approved by FDA have information on
pharmacogenetic biomarkers in the labelling.
Examples of individualized / tailored therapies
 Metastatic breast cancer
• Drug: trastuzumab (Herceptin®)
• Biomarker: HER2 positivity of tumor
 Duchenne muscular dystrophy
• exon skipping
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Source: FDA homepage
Market potential
In 2012
4 of the 10 top selling pharmaceuticals were biologics
What is projected
for 2016?
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Source: Elena Wolff-Holz, European Antibody Congress, 2012; IMS Health Top 20 Global Products 2012
Trends in marketing
authorizations in the US
NOTE: Approvals by the Center for Biologics Evaluation and Research (CBER) are not included
in this drug count.
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Mullard, Nature Reviews Drug Discovery, 2014
Trends in marketing
authorizations in the US / 2
NOTE: Approvals by the CBER are included in the figures.
Dowden et al., Clinical Pharmacology & Therapeutics, 2013
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Costs and risks associated to
biotech drug development
 Approx. $ 1 billion out-of-pocket money for 1 new drug.
≈
 Biotech product development is somewhat more expensive
than small molecule development ($1241 vs $899
million).
Golden Gate bridge, San Francisco
 Average research success rate for clinical development: 1:6.
 Biotech products are somewhat better in terms of pre-market success
rate compared to small molecules.
 Approx. 8-12 years of research for 1 new drug.
DiMasi et al., Manage. Decis. Econ. 2007; DiMasi et al., Nature, 2010; goldengatebridge.org (USD currency in 2003)
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Recent metrics on biotech
Research & Development
From 2001 to 2012 (11 years)…
 the number of biotech products in clinical development grew 155%, from
355 to 907.
 financing of biotech research increased 10-fold, from $10.5 billion to $103
billion.
 worldwide growth in biotechnology product sales grew 353%, from $36 billion to
$163 billion.
In 2012,
the 21 largest pharmaceutical companies had 429 biotech products in
clinical development, of which 58% were monoclonal antibody products.
CSDD, Impact Report, November/December 2013
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Biotechnology in medicine
Milestones in medical biotechnology
Achievements, paradigm shifts
Current trends and research
Conclusions
49
Conclusions
• Some diseases can now be cured / controlled effectively
with biologics, and become part of the standard of care.
• Biologics are important tools of targeted therapy and
help to fulfil the principles of personalized medicine.
• Intensive research is ongoing for new biotech therapies.
• Hungary is strong in the research and production of
biotechnological treatments.
• Further innovative products are expected in the future
especially in the field of the regenerative medicine.
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Thank you for your
attention!
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Questions
& Answers
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