PPT

Report
Passive Immunisierung und
Antikörpertherapien
Konteptvorlesung 3
Themenblock 8
Beda M. Stadler
Institut für Immunologie
KV 8.3
Diese Folien stehen auch als PPT Files zum download bereit unter:
http://www.iib.unibe.ch/wiki/?p=teaching/medical_students&l=de
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Protein Pharmaceutika 2011
• 134 FDA-zugelassene
biotechnologisch hergestellte
(Proteine/Peptide) Medikamente
• Protein Pharmaceutika kosten
derzeit ca. $50 Milliarden/Jahr
2
3
Top Ten Monoclonal Antibodies
2009/2010
Sales $ billion
2009 / 2010
Generic Name
Brands®
Indications
Infliximab
Remicade
RA,UC,CD,Ps,PsA,AS
6.9
8.0
Bevacizumab
Avastin
Colon cancer
5.9
6.8
Rituximab
Rituxan
NHL,RA
5.8
6.7
Adalimumab
Humira
RA,Ps,JIA,PsA,AS,CD
5.4
6.5
Trastuzumab
Herceptin
Breast Cancer
5.0
5.5
Cetuximab
Erbitux
Colon, head and neck cancer
2.5
3.2
Ranibizumab
Lucentis
Wet macular degeneration
2.4
3.1
Natalizumab
Tysabri
Multiple sclerosis
Omalizumab
Xolair
Allergic Asthma
0.9
1.1
Palivizumab
Synagis
RSV
1.1
1.0
1.0
1.7
Thearapeutische Antikörper auch zu teuer für uns?
Übersicht / Lernziele
• Antikörper Produktion einst
• Passive Immuntherapie einst
• Monoklonale /
rekombinante Antikörper
• Antikörper als Therapie für:
• Immunsuppression
• Krebs Therapie
• Immunmodulation
• Anti-Adhäsionstherapien
Paul Ehrlich vor 100 Jahren
• Rückblick und Zukunft
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Passive Immuntherapie einst
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Passive Immuntherapie mit
polyklonalen Antikörpern
Krankheit
Therapeutikum
Schwarze Spinnenbiss
Pferd Antivenin
Botulismus
Pferd Antitoxin
Diphtherie
Pferd Antitoxin
Hepatitis A and B
Multispender human Gammaglobulin
Masern
Multispender human Gammaglobulin
Tollwut
Multispender human Gammaglobulin
Schlangenbiss
Pferd Antivenin
Tetanus
Multispender human Gammaglobulin oder
Pferd Antitoxin
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Monoklonale Antikörper
Hybridoma Technologie
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Therapeutische Maus Antikörper
Probleme
• Immunogenizität
(humane anti-Maus AK, HAMA)
• keine Effektor Mechanismen
(Maus Fc)
• Unerwartete Toxizität
(Reaktivitiät mit normalem Gewebe,
Kreuzreaktion mit anderen Antigenen)
• Verlust von Tumorantigen
(z.B. anti-idiotyp Behandlung von
Lymphomen)
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Immunogenicität von Infliximab
(chimärer monokl. Ak gegen TNFα)
•
61% of patients had
detectable antibodies
after the fifth infusion
of Infliximab
•
Incidence of the
formation of
antibodies to
Infliximab is reduced
with immunosuppressive therapy
Human anti-mouse Ab
Human/mouse chimeric Ab
(Infliximab)
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Diversity of Antibodies is Generated by Gene Rearrangement
Dank Wissen um Grundlagen
rekombinante Antikörper
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Die Domänenstruktur hilft zum
Umbau von Antikörpern
scFv
IgG
(a) Immunoglobulin G
Fab
CL
VL
VH
Hinge
CH
Fv
VL
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VH
CH2
CH3
Fc
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Humanisierung von Maus
Antikörpern
Maus
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Chimäre
Humanisiert
Human
13
Ezzell, Scientific American
Therapeutische Antikörper
Generic name
Trade name
Target
Type
Muromonab
Orthoclone OKT3
CD3
Murine
Abciximab
ReoPro
Platelet GP IIb/IIIa
Murine
Rituximab
Daclizumab
Rituxan
Zenapax
CD20
Chimeric
Humanized
Basiliximab
Palivizumab
Infliximab
Trastuzumab
Gemtuzumab
Alemtuzumab
Simulect
Synagis
Remicade
Herceptin
Mylotarg
Campath-1H
Endrecolomab
IL2Ra
IL2R
RSV
TNFa
Her2/neu/ErbB2
CD33
CD52
Chimeric
Humanized
Chimeric
Humanized
Humanized
Humanized
Panorex
17A-1
Humanized
Ibritumomab tiuxetan
Zevalin
CD20
Chimeric
Adalimumab
Humira
Human
Omalizumab
Xolair
TNFa
IgE
Centuximab
Erbitux
EGFR/ErbB1/Her1
Humanized
Bevacisumab
Avastin
VEGF
Humanized
Efalizumab
Raptiva
CD11a
Humanized
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Humanized
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Mono- & plurivalente Ak-Fragmente
(a) Monovalent antibody fragments
Fab
Fab
Fv
Fv
scFv
scFv
dsFv
dsFv
VL VH
VL VH
VL VH
VH VL
CL
V
VHH
C H1
(b) Single chain two-domain constructs
scFv
scFv
Ag
Diabody
Diabody
Ag
Triabody
Triabody
L
Tetrabody
Tetrabody
or
Ag
Ag
Ag
Ag
L
L Ag
Ag
Ag
VH
VL
L
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VH
VL
VH
Ag
VL
L
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Transgene Humanisation
humane Antikörper in human Ig transgenenen Mäusen
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Der Wunsch nach humanen AK
Humane Antikörper mittels Repertoire Klonierung
mRNA von
B Zellen
Blut
PCR
of
H&L
chain
genes
mRNA  cDNA
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Antikörper (Fab)
Expression auf
p3 des M13 Phagen
Klonierung
und Rekombination
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Expression von Antikörpern
Antikörper
DNA
Transfektion
Antikörper
Vektor
Zellkultur
Expression
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Industrielle Antikörper Produktion
Flask
Spinner
Bioreaktor
(Bench scale)
Bio Reactor
(Plant)
1mL → 100mL → 1L → 1Lx3 → 12L → 100L → 300L → 2’000L→ 10’000L
Subkultur
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Produktion
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Ersatz von passivem IgG
durch rekombinante anti-RhD
RhD+
Erythrozyten
rekombinante Antikörper
mittels phage display
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Neue Therapeutische Antikörper
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Immunsuppressive Anikörper
(anti-T Zell Antikörper)
• Lymphocyte immune globulin (Atgam®)
• lytic to human thymic lymphocytes; blocks T-cell responding
• Anti-CD3: (OKT3®, muromonab-CD3)
• binds CD3, blocks antigen binding; depletes T-cells
• Anti-CD3: huOKT3
• Humanized version of OKT3
• Anti-CD4: OKT4
• less infusion-related reaction
• Anti-Tac monoclonal antibody (Zenopax®)
• binds IL-2 receptor of activated T-cells causing their inactivation
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Lymphocyte Depleting Antibody
Therapies
Polyclonal
Anti-Thymocyte Globulin
Campath-1H
OKT3
CD52
CD3
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Immunsuppressive Anikörper
(anti B-Zell Antikörper)
• Anti-CD20: Rituxan®
(Rituximab)
• results in mature B-cell depletion
and induces apoptosis (B-cell
mediated vascular rejection) for Bcell Non Hodgkin’s Lymphoma
CD19
DR
slg
CD20
CD22
• anti-CD52: Campath-1H
(Alemtuzumab)
• depletes T and B lymphocytes,
NK cells, monocytes and
macrophages
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Side effects
•
•
•
•
•
•
•
•
•
fever
chills
weakness
headache
nausea
vomiting
diarrhea
low blood pressure
rashes
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Antikörper
Krebs Therapie Strategien
2
NK
Complement
pathway
Tumor cell
5
-Y-90 -Y-90
Anti
Angiogenese
-Y-90
1
ADCC
TNF
Anti-Idiotyp
-Y-90
IL-1
-Y-90
Mac
3
-Drug
-Tox
-Tox
4
-Drug
-Tox
-Tox
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Drug
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Strategie 1
Antikörper vermittelte Zytotoxizität
NK Zelle
Ziel Zelle
Ziel Zelle
Apoptosis via
Induktion von intrazellulären
Signal-Pathways
Antikörper-abhängige
Zytotoxizität
(ADCC)
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Ziel Zelle
Complement-abhängige
Zytotoxizität
(CDC)
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z.B. Herceptin® (Trastuzumab)
normal cell
tumor cell
treatment
Binds HER-2 (human epidermal growth factor receptor 2), a
growth factor receptor found on some tumor cells (some breast
cancers, lymphomas).
Over-expression of HER2 causes increased cell growth and
reproduction.
HER2 protein over-expression affects approximately 25% to
30% of breast cancer patients
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Strategie 1: maligne B-Zellen
Stem
cell
ProB cell
PreB cells
Immature
B cell
Mature Activated
B cell
B cell
Antigen independent
Plasma
cell
Antigen dependent
HLA-DR
TDT
CD19
CD10
CD20
CD22
CD21
CD38
Neoplasias:
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Leukemias from B-cell
Precursors (B-ALL)
B-cell Lymphomas
(NHL, CLL)
Multiple Myeloma
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(B-Zell Lymphome)
Strategie 1
anti idiotypische Antikörper
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Strategie 2: Verschiedene Isotope
Nackter Antikörper
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Radiomarkierter Antikörper
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Isotopen markierte Antikörper
LymphoCide (anti CD22,
found on some B-cell
leukemias.
Tositumomab (Anti-CD20)
Lym-1 (Oncolym). AntiHLA-DR expressed at
high levels on lymphoma
cells.
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Strategie 3: Mylotarg® (anti CD-33)
A conjugate of a
monoclonal antibody that
binds CD33 (cell-surface
molecule expressed by the
cancerous cells in acute
myelogenous leukemia
(AML) but not found on the
normal stem cells needed
to repopulate the bone
marrow) and calicheamicin,
an oligosaccharide that
blocks the binding of
transcription factors
(proteins) to DNA and thus
inhibits transcription.
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Strategie 4: Enzym markierte
Antikörper
Antibody directed enzyme
prodrug therapy
(ADEPT)
4
Antikörper
1
2
3
Prodrug
Enzym
Tumor
antigen
Zytotoxischer
Wirkstoff
Tumorzelle
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Strategie 5: Anti-Angiogenes
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Anti-VEGF: Avastin ®
• Recombinant
humanized
monoclonal antibody
against VEGF;
prevents binding with
its receptor.
• Blocks stimulation of
angiogenesis  stops
tumor growth.
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Immunmodulatorische Antikörper
• Anti-TNF Therapie
•
•
•
•
Infliximab
Etanercept
Humira
etc (Biosimilarika)
• Anti-IgE Therapie
• Xolair
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Anti-TNF-a: Infliximab
Chimeric
monoclonal
antibody
• Binds specifically to TNF-a, neutralizing its activity
• Used for Crohn’s disease and rheumatoid arthritis
• Side-effect: can convert a latent case of
tuberculosis into active disease
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Infliximab: Mechanism of Action
Binds and neutralizes soluble and membrane bound TNFa
TNFa
TNFR2
Nucleus
No Signal
transcription
NFB
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Other Postulated Mechanisms of
Action…
Antibody dependent cell-mediated cytotoxicity (ADCC)
Lysis of TNFa-expressing cells through complement (C’) activation
Complement
Activation
C’
Complement
Receptor
Phagocyte
T Cell
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Phagocytosis
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TNF-a: Rezeptor oder Antikörper
Infliximab
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Etanercept
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Biosimilarika: z.B. anti-TNF
Generic
Name
Brand name
(source)
Molecular
Derivation
Remicade®
Human/mouse
chimeric anti-human
TNF monoclonal Ab
Approved in US &
Europe for CD & RA
Etanercept
Enbrel®
Human recombinant
p75 (TNF receptor
fusion protein)
Good for RA
CDP-571
Humicade™
Humanised antihuman TNF mAb
Trials ongoing for CD &
ulcerative colitis
CDP-870
N/A(Celltech)
Humanised anti-TNF
mAb fragment
Phase III trials ongoing
Adalimumab
Humira®
Human anti-human
TNF mAb
Approved for RA
Infliximab
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Development
status
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Omalizumab
Reduces IgE and FcεRI Levels
Omalizumab
(Xolair®)
↓ free IgE
↓ FcεRI
↓ Degranulation
↓ Inflammation
↓ Symptoms
anti-IgE
FcεRI
B
MC
IgE
Holgate S. et al., J Allergy Clin Immunol 2005 vol. 115 (3) pp. 459-65
Mediator
Release
Allergic
Inflammation
Anti-IgE: Omalizumab (Xolair®)
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Anti Adhäsionstherapie
• Efalizumab (Raptiva®)
• Anti-CD11a (subunit of LFA-1) inhibits
activation of T cells
• Alefacept (Amevive®)
• binds to CD2 on memory effector T
lymphocytes, inhibiting their activation
• Abciximab (Reopro®)
• Anti-Gp IIb/IIIa
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Efalizumab (Raptiva)
Interactions between
leukocyte-function
associated antigen type 1
(LFA-1) and intercellular
adhesion molecules are
important in the
pathogenesis of psoriasis.
Efalizumab, a humanized
monoclonal antibody, binds
to the subunit (CD11a) of
LFA-1 and inhibits the
activation of T cells
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Alefacept (Amevive®)
Anti-CD2
CD2
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Abciximab (Reopro®)
Anti-GpIIb/IIIa
Inhibits the clumping of platelets
by binding the receptors on their
surface that normally are linked
by fibrinogen.
Helpful in preventing reclogging of the coronary
arteries in patients who have undergone
angioplasty.
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Ausblick: mehr Antikörper für…
• Transplantation
• Knochenmark, Niere, Leber und Pancreas Insel Zellen.
• Krebs
• Metastatic Brust Krebs, CLL, non-Hodgkin’s Lymphome.
• Autoimmunkrankheiten
• Rheumatoide Arthritis, Psoriasis, Multiple Sclerosis,
Crohn’s Krankheit.
• Infektionskrankheiten
• Respiratory syncytial virus, Cytomegalovirus,
Septikämien.
• Allergien:
• Mehr als 150 weitere Ak in der Pipeline…
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