medical biotechnology - an overview maria judit molnar institute of medical genomics and rare...

Medical biotechnology- an overview

Maria Judit Molnar

Institute of Medical Genomics and Rare Diseases

Semmelweis University

Budapest, Hungary

Conclusions

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Biotechnology in medicine

Milestones in medical biotechnology

Achievements, paradigm shifts

Current trends and research

Biotechnology in medicine

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Biologic medicinal product

Biologicalproduct Biologic

Biologicaldrug

Biotech drug

Biotech

product

=

Definition of biologics

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„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-chemical-biological testing, together with the production process and its control.”

COMMISSION DIRECTIVE 2003/63/EC, 3.2.1.1. b,

Types of biologics

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Biologics

Advanced therapymedicinal products

Somatic cell therapy

Somatic cell therapy

Gene therapyGene therapy

Tissue engineeringTissue engineering

VaccinesVaccines

Blood or blood componentsBlood or blood components

Allergenic productsAllergenic products

Recombinant DNAproducts

Therapeutic proteinsTherapeutic proteins

Monoclonal antibodiesMonoclonal antibodies

Fusion proteinsFusion proteins

XenotransplantationXenotransplantation

COMMISSION DIRECTIVE 2003/63/EC, 3.2.1.1. b,

Biologics vs. Small molecule drugs

6Source: Cecil Nick, TOPRA 2010

Biologics vs. Small molecule drugs

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IN SUMMARY, biologics are protein-like macromolecules. Potential risk: immunogenicity Parenteral administration

Small molecule drugs Biologics

Production Chemical synthesis Extracted from biological source or recombinant DNA techology

Molecular weight < 10 kDa > 10 kDa

Administration route Mainly oral route Mainly parenteral route

Immunogenicity Not an issue Could elicit immunological response

Follow-up molecules Generics Biosimilars

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Milestones in medical biotechnology

Conclusions

Biotechnology in medicine

Achievements, paradigm shifts

Current trends and research

Major milestones in medical biotechnology

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1855

1973

1975

1977

1982

1986

2009

2012

Escherichia coli (E. coli) bacterium discovered (later becomes a major tool for biotechnology).

Genentech scientists and their collaborators produce the first human protein (somatostatin) in a bacterium (E. coli).

Eli Lilly and Company markets recombinant human insulin - the first such product on the market.

Cohen and Boyer develop genetic engineering techniques to "cut and paste" DNA and reproduce the new DNA in bacteria.

First therapeutic monoclonal antibody approved by FDA (muromonab for organ transplant rejection prevention).

Köhler and Milstein: hybridoma technology (Nobel prize 1984).

First tissue engineered product approved in the EU (ChondroCelect).

First gene therapy approved in the EU (Glybera).

Recombinant DNA products

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CLONED CELLS WORK LIKE A FACTORY,

PRODUCING THE DESIRED PROTEIN BESIDES THEIR

OWN PROTEINS.E. coli

Recombinant DNA products manufacturing process

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Multi-step manufacturing process

1. Cell line genetic cloning and selection2. Upscale (= fermentation)3. Downscale (= purification)4. Formulation5. Fill and finish

Cell lines•Prokaryotic (e.g. E. coli)•Eukaryotic (e.g. Chinese Hamster Ovary)

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Milestones in medical biotechnology

Conclusions

Biotechnology in medicine

Achievements, paradigm shifts

Current trends and research

Important therapeutic fields of recombinant DNA products

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Endocrinology

Cardiology

Ophtalmology

Infectious diseases

Transplantation medicine Haematology

Intensive care

Asthma

Oncology

Inflammatory bowel diseases

Neurology

Psoriasis

RheumatologyAutoimmune diseases

ENDOCRINOLOGYInsulin

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From the 1920’sinsulin was derived from porcine / cattle pancreas for therapeutic use (organotherapy)

1982first recombinant insulin on the European market

2014multiple recombinant insulin products on the market

Rapid-actingShort-actingIntermediate-actingLong-actingPre-mixed

ENZYMESGaucher’s disease and

imiglucerase

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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

Pompe Disease

Muscle weakness, atrophyAlpha glycosidase deficiencyAutosomal recessiv

Source: International Society for Mannosidosis & Related Diseases

Monoclonal antibodies(abbreviated as mAbs)

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„…like a key in a keyhole”

TARGETED TARGETED THERAPYTHERAPY

„bind to a corresponding antigen in a highly specific manner…”

Schneider, Curr Pharm Biotechnol., 2008

Monoclonal antibodiesmechanism of action

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antagonism apoptosis CDC ADCC transporter

Complement dependent cytotoxicity

Antigen dependent cellular cytotoxicity

Lutterotti & Martin, Lancet Neurol, 2008

ONCOLOGYTrastuzumab

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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

Source: Herceptin®, European Public Assessment Report

ONCOLOGY + AUTOIMMUNE DISEASE

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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

AUTOIMMUNE DISEASE

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Etanercept• fusion protein• Fc part of a mAb + 2 pieces of TNF-receptors• targeting TNF-alfa

TNF-alfa• cytokine• plays important role in inflammatory processes

Source: Enbrel®, European Public Assessment Report

Indications of etanercept• rheumatoid arthritis• juvenile idiopathic arthritis• ankylosing spondylitis• psoriatic arthritis• plaque psoriasis• paediatric plaque psoriasis

Vaccines

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Prophylactic vaccines against infectious diseases e.g. Hepatitis B vaccination

Therapeutic vaccines

e.g. cancer vaccines intensive research is ongoing

Production derived from natural source recombinant DNA technology

Blood and blood components

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For the treatment of anaemia thrombocytopenia clotting deficiencies

Blood components red blood cells white blood cells plasma clotting factors platelets

Whole blood

Allergenic products

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Application

used for specific immunotherapy (SIT)

in vivo diagnosis of immunoglobulin E (IgE)-mediated allergic diseases

E.g. Prick skin allergy test

Gene therapy

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• First gene therapy approved in EU• Recurrent pancreatitis (lipoprotein lipase deficiency)• Vector: adeno-associated virus

Source: European Medicines Agency homepage

„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”

The history of the gene therapyThe history of the gene therapy

1977 - A gene was successfully delivered into mammalian cells1990 - First human gene therapy was approved: SCID1999 - J. Gelsinger with OTC deficiency died from organ failure

after gene therapy2000 - 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 envelope2009 - 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 The number of gene therapeutical clinical trials

2012 N:3398

2006 N:1064

Phase I N:1023

Phase I/I N:342

Phase II N:1237

Phase II/III N:214

Phase III N:562

General applications ofGeneral applications of therapeutictherapeutic gene transfergene 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

23.04.19.

Vectors

Viral Nonviral

PlasmidArteficial Chr

LiposomesDNAsomes

Nanoparticlees

RetroHerpesAdenoAAVLenti

Strategies

Gene delivery

Molecular therapiesMolecular 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

TheThe Kaplitt Kaplitt ExperimentExperiment

Tp Tp “calm down” the overactive neurons of the subthalamic nucleus which “calm down” the overactive neurons of the subthalamic nucleus which gives rise to signals that cause bradykinesia and tremor in Parkinson’s gives rise to signals that cause bradykinesia and tremor in Parkinson’s

diseasedisease

• 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 SKIPPINGEXON SKIPPING

Exon skipping in Duchenne MDExon skipping in Duchenne MD13% of DMD patients: correct deletions of 50, 52, 45-50, 48-50, 49-50exons13% of DMD patients: correct deletions of 50, 52, 45-50, 48-50, 49-50exons

Progress in AON exon skipping therapy in DMDProgress in AON exon skipping therapy in DMD

TimelinesTimelines 2007 2008 2009 2010 2011 2012 20132007 2008 2009 2010 2011 2012 2013

PRO051Prosensa/GSK

2OME AON

EteplirsenAVI

PMO AON

Ph IIm.

Ph I/IISystemic adm, iv, weekly

Ph I/IIStudy extension.

Ph INon ambulant

Ph IIDosing

Ph IIIEfficacy

Ph IIm.

Ph I/IISyst. admin. sc, 3 w

Ph I/IIDosing

P15 P17 P18

P19

Pre

Post

Post Pre Post Pre Post Pre Post Pre

Results: Lancet 2011 Aug 13; 378(9791):595-605

P15 P17 P18

• 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

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• approved in the EU (2013)• autologous peripheral blood mononuclear cells activated with PAP-GM-CSF

Sources: European Medicines Agency homepage; Barar & Omidi, BioImpacts, 2012

„contain cells or tissues that have been manipulated to change their biological characteristics. They can be used to cure, diagnose or prevent diseases”

Stem cells

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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

„cells with self-renewing capacity i.e. the capability to generate daughter cells

and multi-lineage differentiation capacity. Stem cells are capable of proliferation as stem cells in an undifferentiated form”

Stem cells

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e.g. ReNeuron’s ReN001

Phase 1 trial is ongoing, phase 2 trial application submitted

For the treatment of ischemic stroke

Potential applications of stem cells

metabolic, degenerative and inflammatory diseases

repair and regeneration of damaged or lost tissues

treatment of cancer

Source: EMA/CAT/571134/2009; ReNeuron homepage

Umbilical cord stem cell preservation

Tissue engineered products

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ChondroCelecto first advanced therapy medicinal product in the EU in 2009

MACI o matrix autologous chondrocyte implantationo approved in EU in 2013

Source: European Medicines Agency homepage

„contain cells or tissues that have been modified so they can be used to

repair, regenerate or replace tissue”

Regenerative medicine

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3D printing

Bone grafts

Blood vessels

Ear printing

Printing of a kidney

Liver printing with hepatocytes (awaited for 2014)

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Milestones in medical biotechnology

Conclusions

Biotechnology in medicine

Achievements, paradigm shifts

Current trends and research

Personalized medicine

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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

"the right patient with the right drug at the right dose at the right time."

Source: FDA homepage

Market potential

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In 20124 of the 10 top selling pharmaceuticals were biologics

What is projectedfor 2016?

Source: Elena Wolff-Holz, European Antibody Congress, 2012; IMS Health Top 20 Global Products 2012

Trends in marketing authorizations in the US

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NOTE: Approvals by the Center for Biologics Evaluation and Research (CBER) are not included in this drug count.

Mullard, Nature Reviews Drug Discovery, 2014

Trends in marketing authorizations in the US / 2

46Dowden et al., Clinical Pharmacology & Therapeutics, 2013

NOTE: Approvals by the CBER are included in the figures.

Costs and risks associated to biotech drug development

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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).

DiMasi et al., Manage. Decis. Econ. 2007; DiMasi et al., Nature, 2010; goldengatebridge.org (USD currency in 2003)

≈

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.

Golden Gate bridge, San Francisco

Approx. 8-12 years of research for 1 new drug.

Recent metrics on biotech Research & Development

48CSDD, Impact Report, November/December 2013

In 2012, the 21 largest pharmaceutical companies had 429 biotech products in clinical development, of which 58% were monoclonal antibody products.

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.

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Milestones in medical biotechnology

Conclusions

Biotechnology in medicine

Achievements, paradigm shifts

Current trends and research

Conclusions

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• 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