1. Tumor Therapy with Monoclonal Antibodies
Gerhard Moldenhauer

2. Innate and Adaptive Immunity

3. Lymphocyte Activation

4. Effector Functions of Antibodies

5. Model of Secreted IgG

6. Structure of an IgG Antibody
Original by Dr. Mike Clark

7. Monoclonal Antibodies
Generation of
Monoclonal Antibodies
G. Köhler and C. Milstein
1975

8. Characteristics of Monoclonal Antibodies

9. Mechanisms Operating with Therapeutic Antibodies

10. Therapeutic Effects of Monoclonal Antibodies

11. Strategies for Therapeutic Application of Monoclonal Antibodies

12. Patient Suffering from NHL

13. Immunocytology of Bone Marrow Smear
from a Patient with NHL

14. Non-Hodgkin‘s lymphomas

15. Target Antigens on Malignant B-cells
HLA-DR
TDT
CD19
CD10
CD20
CD22
CD21
CD38
Stem
cell
Pro-
B cell
Pre-
B cells
Immature
Mature
Activated
Plasma
Neoplasias:
Leukemias from B-cell
Precursors (B-ALL)
B-cell Lymphomas
(NHL, CLL)
Multiple Myeloma
Antigen independent
Antigen dependent

16. First Report on Treatment of a Cancer Patient with Monoclonal Antibody

17. Monoclonal Antibody Therapy - Problems

18. Domain Structure of Immunoglobulins
Fab Fv Fc
Hinge VL VH CL CH
CH2
CH3
Domain Structure of Immunoglobulins
scFv
IgG

19. Antibodies for Therapeutic Application
Human
Murine
Chimeric
Humanized
Antibodies for Therapeutic Application
scFv
Fragment
Bispecific

20. Rituximab

21. Rituximab

22. Strategies for Therapeutic Application of Monoclonal Antibodies

23. Kinase Growth Factor Pathway
Cell membrane
Ligand binding
Activated receptor Y P
Proliferation
Migration
Tumour growth and metastases
Survival
Signal transduction
Tyrosine kinase receptor
Tyrosine
kinase
domain
Original by Dr. Axel Ullrich

28. Effects of Trastuzumab (Herceptin) on Breast Cancer Cells

29. EGFR signaling
The EGFR is activated by growth factors (e.g. epidermal growth factor (EGF) and transforming growth factor- (TGF-)).
EGFR-activation leads to the building of either receptor homo- or hetero-dimers.
Receptor dimerization initiates an intracellular signaling cascade, gene activation and the stimulation of cell cycle progression.
- The epidermal growth factor receptor (EGFR) is a transmembrane receptor with an intracellular tyrosine kinase domain, which is activated by growth factors. The main activating ligands are epidermal growth factor (EGF) and transforming growth factor- (TGF- ) [1].
- EGFR is a protooncogene and its activation plays a key role in the development of many cancers. Activation occurs because of gene amplification, gene mutation and/or autocrine stimulation due to the overproduction of the EGFR ligands (eg EGF and TGF- ). Activation leads to the pairing of receptors with either another EGFR (homodimers) or another member of the EGFR family (heterodimers) [1].
- Receptor dimerization initiates an intracellular signaling cascade, gene activation and the stimulation of cell cycle progression [1].
1. Baselga J. Eur J Cancer 2001; 37 Suppl 4:S16–S22.

30. The importance of EGFR as a target
“Evidence for a role for the EGFR in the inhibition and pathogenesis of various cancers has led to the rational design and development of agents that selectively target this receptor.”*
- The EGFR intracellular signaling cascade stimulates not only cell proliferation but also protection from apoptosis, loss of differentiation, angiogenesis, cell migration and metastasis formation (ie all the key processes involved in tumorigenesis) [1].
- EGFR is expressed in a high proportion of solid tumors, in particular head and neck, lung and colorectal cancer [2]. Expression has been correlated with disease progression [3]. Many studies have shown that EGFR expression can be an adverse prognostic factor for cancer treatment outcome [2].
- ‘Evidence for a role for the EGFR in the inhibition and pathogenesis of various cancers has led to the rational design and development of agents that selectively target this receptor,’ Baselga 2002 [3].
1. Baselga J. Eur J Cancer 2001; 37 Suppl 4:S16–S22.
2. Nicholson RI, Gee JMW, Harper ME. Eur J Cancer 2001; 37 Suppl 4:S9–S15.
3. Baselga J. The Oncologist 2002; 7 Suppl 4:2–8.
* Baselga 2002

31. Erbitux® (cetuximab)
Erbitux® (cetuximab) is an IgG1 MAb targeting the EGFR
Binding blocks EGFR signaling and inhibits proliferation, angio-genesis and metastasis, and stimulates apoptosis and differentiation
The main toxicity is an acne-like rash that generally improves during treatment, and usually does not preclude continued treatment
- Erbitux® (cetuximab) is an IgG1 MAb that targets EGFR with a high specificity and a higher affinity than either of the two main activating growth factors (EGF and TGF-) [1].
- The binding of Erbitux® to EGFR blocks ligand-induced EGFR signaling and results in the inhibition of proliferation, - stimulation of apoptosis, inhibition of angiogenesis, inhibition of de-differentiation and prevention of metastasis formation [1]. Being an IgG1 monoclonal antibody Erbitux® has been shown to mediate antibody-dependent cellular cytotoxicity (ADCC) in vitro [2]. Therefore, in addition to its inhibitory function on receptor signaling, patients with EGFR-expressing tumors may also benefit from this immune stimulatory effect of Erbitux®.
- The antibody has a long half-life of around 3-4 days, making it suitable for convenient weekly administration. Erbitux® is administered 400 mg/m2 for the 1st infusion followed by 250 mg/m2 weekly.
- Erbitux® is well tolerated, lacks the hematologic toxicity [3] commonly associated with a number of anticancer agents, and does not exacerbate the toxicity of the investigated chemotherapeutic drugs [4]. The main side effect is an acne-like rash which generally improves during treatment and usually does not preclude continued treatment [5].
1. Baselga J. Eur J Cancer 2001; 37 Suppl 4:S16–S22.
2. Naramura M, Gillies SD, Mendelsohn J, et al. Cancer Immunol Immunother 1993; 37:
3. Mendelsohn J. J Clin Oncol 2002; 20 Suppl 18:1S–13S.
4. O’Dwyer PJ, Benson AB. Semin Oncol 2002; 29 Suppl 14:10–17.
5. Needle MN. Semin Oncol 2002; 29 Suppl 14:55–60.

32. Deborah Schrag : The Price Tag on Progress N Engl J Med, July 22, 2004
“Just how much are we as a society willing to spend on the treatment of advanced cancer ?“

33. Strategies for Therapeutic Application of Monoclonal Antibodies

34. Inhibition of Angiogenesis

41. Strategies for Therapeutic Application of Monoclonal Antibodies

42. Internal Radiation by Radiolabeled mAb

43. Immunoscintigraphy with 131I-labeled mAb

44. RIT: Clinical Trials for NHL
* myeloablative (autologous stem cell rescue)
Antibody
Antigen
Nuclide
Patients n
Response
% (CR)
Witzig et al. 2002
Ibritumomab
(Zevalin®)
CD20
90Y 54
74 (15)
Kaminski et al. 2001
Tositumomab
(BexxarTM)
131I 60
65 (20)
De Nardo et al. 1998
Lym-1
HLA-DR 51
75 (20)
Juweid et al. 1999
HLL2
CD22
131I, 90Y 22
18 (5)
Press et al * B1 29
86 (79)

45. Strategies for Therapeutic Application of Monoclonal Antibodies

46. Immunotoxin

47. Recombinant Immunotoxin Fusion Protein
tumor
cell
normal
ligand
A-chain
B-chain
immunotoxin
facilitates
delivery
into cytosol
binds to many cells,
must be blocked
or removed
inhibits
protein
synthesis
mAb,
scFv,
cytokine
receptor
cytotoxic
domain
translocation
binding
toxin moiety
Recombinant Immunotoxin Fusion Protein
Original by Dr. D. Vallera

48. Strategies for Therapeutic Application of Monoclonal Antibodies

49. Monoclonal anti-Idiotype Antibodies

50. Strategies for Therapeutic Application of Monoclonal Antibodies

51. Cytotoxic T-cell Targeting
Kill
EGF-R
MUC-1
cytotoxic
T lymphocyte
carcinoma cell
CD3
Ep-CAM
bispecific antibody
HEA125xCD3
Cytotoxic T-cell Targeting

52. Ovarian Carcinoma Stained for Ep-CAM Expression

53. Selection of QuadromasX
Hybridoma A (HGPRT-)
Hybridoma B (Iodoacetamide)
HAT
Hybrid-Hybridoma

54. Antibody Mixture Produced by Hybrid-Hybridomas

55. Ovarian Cancer
¨ Leading cause of death among gynecological malignancies
¨Therapy consists of cytoreductive surgery and combination chemotherapy
¨ Prognosis of advanced disease is very poor (5 year survival at stages III and IV less than 20%)
=> Early spread of carcinoma cells from ovaries into the peritoneal cavity giving rise to multiple metastases
=> Formation of malignant ascites leading to severe symptoms (anorexia, dyspnea, obstruction of the gastrointestinal tract)

56. Clinical Trial of Intraperitoneal BsAb Therapy
in Patients with Advanced Ovarian Cancer

57. Changes in Ascites Volume during bsAb Therapy
Patient 2
All patients
Ascites production
lowest value
during therapy
Patient 1 ,
020
263 2
135 3
218 4
190 5
533 6
290 7
571
350 8
000 9
200 10
320
before therapy
Ascites volume in ml/day
HEA125xOKT3 applications
800
4000
ascites ml/d
700
3500
ascites volume
CA125 U/ml
600
3000
CA125 serum level
500
2500
400
2000
300
1500
200
1000
100
500
8 month of follow up
Collaboration with the Department of Obstetrics and Gynecology at the University of Heidelberg
(Dr. A. Marmé, Prof. G. Bastert)

58. Eighteen Therapeutic Antibodies Approved by the FDA

59. Paul Ehrlich (1854-1915) Finally, the dreams of Paul Ehrlich
who considered antibodies
as magic bullets have become
reality.
Monoclonal antibodies have
established themselves as the most
important and rapidly expanding
class of drugs in oncology.
Paul Ehrlich
( )