Applications of Biotechnological Processes Antibody Production.
BIOTECHNOLOGICAL APPLICATIONS 1Therapeutic antibodies 6_humanization
-
Upload
free-education-for-all -
Category
Education
-
view
66 -
download
1
Transcript of BIOTECHNOLOGICAL APPLICATIONS 1Therapeutic antibodies 6_humanization
Is Ofatumumab Better than Rituximab
Ofatumumab is a fully human antibody produced in transgenic mice immunized with CD20
bull UK-based GlaxoSmithKline and Danish biotech firm Genmab have reported results from a Phase III trial (ORCHARRD) of ofatumumab (Arzerra) plus chemotherapy versus rituximab plus chemotherapy to treat patients with relapsed or refractory diffuse large B-cell lymphoma (DLBCL)
bull The trial did not meet its primary endpoint as there was no statistically significant difference in progression free survival (PFS) between the treatment arms
bull According to the firms there were no differences in adverse events (AEs) leading to treatment discontinuation Grade gt3 AEs severe adverse events (SAEs) or fatal SAEs between the treatment arms
bull Based on todays results we are unlikely to move forward with a regulatory filing Winkel said
Allora Is There Anything Our There that can Beat Rituximab in a Head to Head Trial
There are at least three mechanisms by which therapeutic antibodies targeting CD20 destroy their target cells
Polymorphisms in FcγRIIIa Receptor Influence the Therapeutic Activity of Retuximab
bull FCGR3A gene encodes FcγRIIIa receptor
bull Fcγ receptors are found on immune effector cells that mediate Antibody-Dependent Cell Cytotoxicity (ADCC)
bull FcγRIIIa is polymorphic at position 158 ndash Val or Phe
56
35 P = 02
49 patients with B cell follicular lymphoma
Cartron et al (2002) Blood
Finding Statistical Differences by Looking at Data in a Different Way
CRu complete remission
unconfirmed
Interaction of Immunoglobulins with FcγRIII Receptors
polymorphic position 158
AgAg
bivalent
Cold turkey ndash describes the actions of a person who abruptly gives up a habit or addiction rather than gradually easing the process
Immunoglobulins are Subject to N-Linked Glycosylation
Glycoproteins
N-linked glycosylation ndash oligosaccharides linked to Asn residues in the sequence context
Asn-X-Ser or Asn-X-Thr
Proteins are initially modified by a 14 residues core oligosaccharide
Dolichol is a lipid
CH2
C ONSugar
N-glycosidic bond
Hunusual high energy phosphate
ester linkage provides energy for forming N-glycosidic bond
Cotranslational Addition of the Core Oligosaccharide
oligosaccharide transferase
signal peptide
cytoplasm
rough ER membrane
ER lumen
complex
high mannose
Asn-X-SerAsn-X-Thr
Processing the Core Oligosaccharide
Glucose
Mannose
N-acetyl glucosamine
Fucose
Galactose
Sialic acid
High mannose class switches from trimming to addition
Complex class switches from trimming to addition
Antibodies are Glycoproteins and Glycosylation can Influence Effector Function
Antibodies are subject to N-linked glycosylation at residue 297 of the Fc region
Modifying Glycosylation Patterns
Biantennary fucosylated complex oligosaccharide
bull 111308914-N-acetylglucosaminyltransferase III (GnT-III)
GluNAc
Man
Fuc
bull 1113089Man-2
Transfect cells with
After glyco-engineering bisected non-fucosylated antibody
Kinetic Constants for the Binding of Antibodies with Modified Carbohydrates to FcγRIIIa
20 of patients are homozygous Valine
35 of patients are homozygous Phenyalanine
Biacor Traditional equilibrium
binding assay
bull GA101 with native glycosylation pattern binds much better to polymorphic variant with Val
bull GA101 with modified glycosylation pattern binds to either polymorphic variant with roughly equal affinity
Cell Death Assays Annexin Staining
Phosphatidylserine (PS) normally found on cytoplasmic surface of plasma membrane
During intermediate stages of apoptosis PS is translocated to the outer surface of the plasma membrane
Externalization of PS makes it available for binding to fluorescent PS-binding proteins (Annexins)
Added one additional mutation (V for L) in the hinge region
Selected on the basis of ADCC rather than affinity
GA101 Does Not Cause CD20 to Aggregate in Lipid Rafts Like Rituximab
Antibodies to CD20 fall into two classes
bull Class 1 ndash includes Rituximab causes CD20 to aggregate in lipid rafts
bull Class 2 ndash no aggregation fewer molecules bound per cell
G R
Rituximab is more Effective than GA101 at Promoting Complement-Dependent Cell Lysis
rituximab
GA101
GA101 is more Effective at ADCC and B Cell Depletion than Rituximab
GA101
rituximab
GA101 is more Effective than Rituximab in Mouse Xenograph Models of Diffuse B Cell Lymphoma
only rituximab
Rituximab or GA101
GA101 is More Effective than Rituximab in Depleting B Cells in Clonus Monkeys
G R
Of Trumps Rituximab in a Clinical Trial for Chronic lymphocytic Leukemia
One Last Look at Where We would be Without Therapeutic Antibodies
Imatinib and Beyond Protein Kinase Inhibitors as Cancer Therapeutics
Protein Kinases as Drug Targets
Problem
There are over 500 protein kinases encoded by the human genome Can a drug target just one
Cancer Drug Gleevec (Imatinib) Wins Lasker AwardPosted at 343 pm CT on September 14 2009
The shortened Philadelphia chromosome seen in certain leukemias (picture from naturecom)The big science news of the day was the announcement of the Lasker Awards informally thought of as the American version of the Nobel Prize for physiology and medicine This yearrsquos clinical medical research award went to a trio of researchers from Oregon Health amp Science University Sloan-Kettering Cancer Center and drug company Novartis but you could just as easily say it was awarded to a drug the cancer treatment Gleevec (imatinib)
Noyes
sufficient specificity for clinical benefit
Imatinib and Related Drugs Target Philadelphia Chromosome Positive CML
bull BCR-ABL fusion creates a dys-regulated protein kinase that drives the proliferation of CML clones
protein kinase active site
bind ATP
bind target protein substrate
imatinib
specific kinase inhibition
A priori targeting the protein substrate binding site might be expected to provide more specificity buthelliphelliphelliphelliphelliphelliphellip
The Imatinib Experience
bull 82 of patients achieve a complete cytogenetic response (65-85)bull majority achieve a major molecular response (40-60)bull 7 year overall survival rate 90 bull event free survival rate 81 (65-85)
Positives
results from the International Randomized Study of Interferon Versus STI571 (IRIS) trial 7-year follow up
Blood (2008) 112 186
various studies from 2003-2008
bull Imatinib however does not destroy the cancer stem cell clone
bull If you remove imatinib CML returns
bull Imatinib is the perfect drug because once you are on it you are on it forever
If you place a cancer cell under selective pressure for long enough without killing it what will happen
- Slide 1
- Slide 2
- Slide 3
- Slide 4
- Slide 5
- Slide 6
- Slide 7
- Slide 8
- Slide 9
- Slide 10
- Slide 11
- Slide 12
- Slide 13
- Slide 14
- Slide 15
- Slide 16
- Slide 17
- Slide 18
- Slide 19
- Slide 20
- Slide 21
- Slide 22
- Slide 23
- Slide 24
-
Allora Is There Anything Our There that can Beat Rituximab in a Head to Head Trial
There are at least three mechanisms by which therapeutic antibodies targeting CD20 destroy their target cells
Polymorphisms in FcγRIIIa Receptor Influence the Therapeutic Activity of Retuximab
bull FCGR3A gene encodes FcγRIIIa receptor
bull Fcγ receptors are found on immune effector cells that mediate Antibody-Dependent Cell Cytotoxicity (ADCC)
bull FcγRIIIa is polymorphic at position 158 ndash Val or Phe
56
35 P = 02
49 patients with B cell follicular lymphoma
Cartron et al (2002) Blood
Finding Statistical Differences by Looking at Data in a Different Way
CRu complete remission
unconfirmed
Interaction of Immunoglobulins with FcγRIII Receptors
polymorphic position 158
AgAg
bivalent
Cold turkey ndash describes the actions of a person who abruptly gives up a habit or addiction rather than gradually easing the process
Immunoglobulins are Subject to N-Linked Glycosylation
Glycoproteins
N-linked glycosylation ndash oligosaccharides linked to Asn residues in the sequence context
Asn-X-Ser or Asn-X-Thr
Proteins are initially modified by a 14 residues core oligosaccharide
Dolichol is a lipid
CH2
C ONSugar
N-glycosidic bond
Hunusual high energy phosphate
ester linkage provides energy for forming N-glycosidic bond
Cotranslational Addition of the Core Oligosaccharide
oligosaccharide transferase
signal peptide
cytoplasm
rough ER membrane
ER lumen
complex
high mannose
Asn-X-SerAsn-X-Thr
Processing the Core Oligosaccharide
Glucose
Mannose
N-acetyl glucosamine
Fucose
Galactose
Sialic acid
High mannose class switches from trimming to addition
Complex class switches from trimming to addition
Antibodies are Glycoproteins and Glycosylation can Influence Effector Function
Antibodies are subject to N-linked glycosylation at residue 297 of the Fc region
Modifying Glycosylation Patterns
Biantennary fucosylated complex oligosaccharide
bull 111308914-N-acetylglucosaminyltransferase III (GnT-III)
GluNAc
Man
Fuc
bull 1113089Man-2
Transfect cells with
After glyco-engineering bisected non-fucosylated antibody
Kinetic Constants for the Binding of Antibodies with Modified Carbohydrates to FcγRIIIa
20 of patients are homozygous Valine
35 of patients are homozygous Phenyalanine
Biacor Traditional equilibrium
binding assay
bull GA101 with native glycosylation pattern binds much better to polymorphic variant with Val
bull GA101 with modified glycosylation pattern binds to either polymorphic variant with roughly equal affinity
Cell Death Assays Annexin Staining
Phosphatidylserine (PS) normally found on cytoplasmic surface of plasma membrane
During intermediate stages of apoptosis PS is translocated to the outer surface of the plasma membrane
Externalization of PS makes it available for binding to fluorescent PS-binding proteins (Annexins)
Added one additional mutation (V for L) in the hinge region
Selected on the basis of ADCC rather than affinity
GA101 Does Not Cause CD20 to Aggregate in Lipid Rafts Like Rituximab
Antibodies to CD20 fall into two classes
bull Class 1 ndash includes Rituximab causes CD20 to aggregate in lipid rafts
bull Class 2 ndash no aggregation fewer molecules bound per cell
G R
Rituximab is more Effective than GA101 at Promoting Complement-Dependent Cell Lysis
rituximab
GA101
GA101 is more Effective at ADCC and B Cell Depletion than Rituximab
GA101
rituximab
GA101 is more Effective than Rituximab in Mouse Xenograph Models of Diffuse B Cell Lymphoma
only rituximab
Rituximab or GA101
GA101 is More Effective than Rituximab in Depleting B Cells in Clonus Monkeys
G R
Of Trumps Rituximab in a Clinical Trial for Chronic lymphocytic Leukemia
One Last Look at Where We would be Without Therapeutic Antibodies
Imatinib and Beyond Protein Kinase Inhibitors as Cancer Therapeutics
Protein Kinases as Drug Targets
Problem
There are over 500 protein kinases encoded by the human genome Can a drug target just one
Cancer Drug Gleevec (Imatinib) Wins Lasker AwardPosted at 343 pm CT on September 14 2009
The shortened Philadelphia chromosome seen in certain leukemias (picture from naturecom)The big science news of the day was the announcement of the Lasker Awards informally thought of as the American version of the Nobel Prize for physiology and medicine This yearrsquos clinical medical research award went to a trio of researchers from Oregon Health amp Science University Sloan-Kettering Cancer Center and drug company Novartis but you could just as easily say it was awarded to a drug the cancer treatment Gleevec (imatinib)
Noyes
sufficient specificity for clinical benefit
Imatinib and Related Drugs Target Philadelphia Chromosome Positive CML
bull BCR-ABL fusion creates a dys-regulated protein kinase that drives the proliferation of CML clones
protein kinase active site
bind ATP
bind target protein substrate
imatinib
specific kinase inhibition
A priori targeting the protein substrate binding site might be expected to provide more specificity buthelliphelliphelliphelliphelliphelliphellip
The Imatinib Experience
bull 82 of patients achieve a complete cytogenetic response (65-85)bull majority achieve a major molecular response (40-60)bull 7 year overall survival rate 90 bull event free survival rate 81 (65-85)
Positives
results from the International Randomized Study of Interferon Versus STI571 (IRIS) trial 7-year follow up
Blood (2008) 112 186
various studies from 2003-2008
bull Imatinib however does not destroy the cancer stem cell clone
bull If you remove imatinib CML returns
bull Imatinib is the perfect drug because once you are on it you are on it forever
If you place a cancer cell under selective pressure for long enough without killing it what will happen
- Slide 1
- Slide 2
- Slide 3
- Slide 4
- Slide 5
- Slide 6
- Slide 7
- Slide 8
- Slide 9
- Slide 10
- Slide 11
- Slide 12
- Slide 13
- Slide 14
- Slide 15
- Slide 16
- Slide 17
- Slide 18
- Slide 19
- Slide 20
- Slide 21
- Slide 22
- Slide 23
- Slide 24
-
Polymorphisms in FcγRIIIa Receptor Influence the Therapeutic Activity of Retuximab
bull FCGR3A gene encodes FcγRIIIa receptor
bull Fcγ receptors are found on immune effector cells that mediate Antibody-Dependent Cell Cytotoxicity (ADCC)
bull FcγRIIIa is polymorphic at position 158 ndash Val or Phe
56
35 P = 02
49 patients with B cell follicular lymphoma
Cartron et al (2002) Blood
Finding Statistical Differences by Looking at Data in a Different Way
CRu complete remission
unconfirmed
Interaction of Immunoglobulins with FcγRIII Receptors
polymorphic position 158
AgAg
bivalent
Cold turkey ndash describes the actions of a person who abruptly gives up a habit or addiction rather than gradually easing the process
Immunoglobulins are Subject to N-Linked Glycosylation
Glycoproteins
N-linked glycosylation ndash oligosaccharides linked to Asn residues in the sequence context
Asn-X-Ser or Asn-X-Thr
Proteins are initially modified by a 14 residues core oligosaccharide
Dolichol is a lipid
CH2
C ONSugar
N-glycosidic bond
Hunusual high energy phosphate
ester linkage provides energy for forming N-glycosidic bond
Cotranslational Addition of the Core Oligosaccharide
oligosaccharide transferase
signal peptide
cytoplasm
rough ER membrane
ER lumen
complex
high mannose
Asn-X-SerAsn-X-Thr
Processing the Core Oligosaccharide
Glucose
Mannose
N-acetyl glucosamine
Fucose
Galactose
Sialic acid
High mannose class switches from trimming to addition
Complex class switches from trimming to addition
Antibodies are Glycoproteins and Glycosylation can Influence Effector Function
Antibodies are subject to N-linked glycosylation at residue 297 of the Fc region
Modifying Glycosylation Patterns
Biantennary fucosylated complex oligosaccharide
bull 111308914-N-acetylglucosaminyltransferase III (GnT-III)
GluNAc
Man
Fuc
bull 1113089Man-2
Transfect cells with
After glyco-engineering bisected non-fucosylated antibody
Kinetic Constants for the Binding of Antibodies with Modified Carbohydrates to FcγRIIIa
20 of patients are homozygous Valine
35 of patients are homozygous Phenyalanine
Biacor Traditional equilibrium
binding assay
bull GA101 with native glycosylation pattern binds much better to polymorphic variant with Val
bull GA101 with modified glycosylation pattern binds to either polymorphic variant with roughly equal affinity
Cell Death Assays Annexin Staining
Phosphatidylserine (PS) normally found on cytoplasmic surface of plasma membrane
During intermediate stages of apoptosis PS is translocated to the outer surface of the plasma membrane
Externalization of PS makes it available for binding to fluorescent PS-binding proteins (Annexins)
Added one additional mutation (V for L) in the hinge region
Selected on the basis of ADCC rather than affinity
GA101 Does Not Cause CD20 to Aggregate in Lipid Rafts Like Rituximab
Antibodies to CD20 fall into two classes
bull Class 1 ndash includes Rituximab causes CD20 to aggregate in lipid rafts
bull Class 2 ndash no aggregation fewer molecules bound per cell
G R
Rituximab is more Effective than GA101 at Promoting Complement-Dependent Cell Lysis
rituximab
GA101
GA101 is more Effective at ADCC and B Cell Depletion than Rituximab
GA101
rituximab
GA101 is more Effective than Rituximab in Mouse Xenograph Models of Diffuse B Cell Lymphoma
only rituximab
Rituximab or GA101
GA101 is More Effective than Rituximab in Depleting B Cells in Clonus Monkeys
G R
Of Trumps Rituximab in a Clinical Trial for Chronic lymphocytic Leukemia
One Last Look at Where We would be Without Therapeutic Antibodies
Imatinib and Beyond Protein Kinase Inhibitors as Cancer Therapeutics
Protein Kinases as Drug Targets
Problem
There are over 500 protein kinases encoded by the human genome Can a drug target just one
Cancer Drug Gleevec (Imatinib) Wins Lasker AwardPosted at 343 pm CT on September 14 2009
The shortened Philadelphia chromosome seen in certain leukemias (picture from naturecom)The big science news of the day was the announcement of the Lasker Awards informally thought of as the American version of the Nobel Prize for physiology and medicine This yearrsquos clinical medical research award went to a trio of researchers from Oregon Health amp Science University Sloan-Kettering Cancer Center and drug company Novartis but you could just as easily say it was awarded to a drug the cancer treatment Gleevec (imatinib)
Noyes
sufficient specificity for clinical benefit
Imatinib and Related Drugs Target Philadelphia Chromosome Positive CML
bull BCR-ABL fusion creates a dys-regulated protein kinase that drives the proliferation of CML clones
protein kinase active site
bind ATP
bind target protein substrate
imatinib
specific kinase inhibition
A priori targeting the protein substrate binding site might be expected to provide more specificity buthelliphelliphelliphelliphelliphelliphellip
The Imatinib Experience
bull 82 of patients achieve a complete cytogenetic response (65-85)bull majority achieve a major molecular response (40-60)bull 7 year overall survival rate 90 bull event free survival rate 81 (65-85)
Positives
results from the International Randomized Study of Interferon Versus STI571 (IRIS) trial 7-year follow up
Blood (2008) 112 186
various studies from 2003-2008
bull Imatinib however does not destroy the cancer stem cell clone
bull If you remove imatinib CML returns
bull Imatinib is the perfect drug because once you are on it you are on it forever
If you place a cancer cell under selective pressure for long enough without killing it what will happen
- Slide 1
- Slide 2
- Slide 3
- Slide 4
- Slide 5
- Slide 6
- Slide 7
- Slide 8
- Slide 9
- Slide 10
- Slide 11
- Slide 12
- Slide 13
- Slide 14
- Slide 15
- Slide 16
- Slide 17
- Slide 18
- Slide 19
- Slide 20
- Slide 21
- Slide 22
- Slide 23
- Slide 24
-
Finding Statistical Differences by Looking at Data in a Different Way
CRu complete remission
unconfirmed
Interaction of Immunoglobulins with FcγRIII Receptors
polymorphic position 158
AgAg
bivalent
Cold turkey ndash describes the actions of a person who abruptly gives up a habit or addiction rather than gradually easing the process
Immunoglobulins are Subject to N-Linked Glycosylation
Glycoproteins
N-linked glycosylation ndash oligosaccharides linked to Asn residues in the sequence context
Asn-X-Ser or Asn-X-Thr
Proteins are initially modified by a 14 residues core oligosaccharide
Dolichol is a lipid
CH2
C ONSugar
N-glycosidic bond
Hunusual high energy phosphate
ester linkage provides energy for forming N-glycosidic bond
Cotranslational Addition of the Core Oligosaccharide
oligosaccharide transferase
signal peptide
cytoplasm
rough ER membrane
ER lumen
complex
high mannose
Asn-X-SerAsn-X-Thr
Processing the Core Oligosaccharide
Glucose
Mannose
N-acetyl glucosamine
Fucose
Galactose
Sialic acid
High mannose class switches from trimming to addition
Complex class switches from trimming to addition
Antibodies are Glycoproteins and Glycosylation can Influence Effector Function
Antibodies are subject to N-linked glycosylation at residue 297 of the Fc region
Modifying Glycosylation Patterns
Biantennary fucosylated complex oligosaccharide
bull 111308914-N-acetylglucosaminyltransferase III (GnT-III)
GluNAc
Man
Fuc
bull 1113089Man-2
Transfect cells with
After glyco-engineering bisected non-fucosylated antibody
Kinetic Constants for the Binding of Antibodies with Modified Carbohydrates to FcγRIIIa
20 of patients are homozygous Valine
35 of patients are homozygous Phenyalanine
Biacor Traditional equilibrium
binding assay
bull GA101 with native glycosylation pattern binds much better to polymorphic variant with Val
bull GA101 with modified glycosylation pattern binds to either polymorphic variant with roughly equal affinity
Cell Death Assays Annexin Staining
Phosphatidylserine (PS) normally found on cytoplasmic surface of plasma membrane
During intermediate stages of apoptosis PS is translocated to the outer surface of the plasma membrane
Externalization of PS makes it available for binding to fluorescent PS-binding proteins (Annexins)
Added one additional mutation (V for L) in the hinge region
Selected on the basis of ADCC rather than affinity
GA101 Does Not Cause CD20 to Aggregate in Lipid Rafts Like Rituximab
Antibodies to CD20 fall into two classes
bull Class 1 ndash includes Rituximab causes CD20 to aggregate in lipid rafts
bull Class 2 ndash no aggregation fewer molecules bound per cell
G R
Rituximab is more Effective than GA101 at Promoting Complement-Dependent Cell Lysis
rituximab
GA101
GA101 is more Effective at ADCC and B Cell Depletion than Rituximab
GA101
rituximab
GA101 is more Effective than Rituximab in Mouse Xenograph Models of Diffuse B Cell Lymphoma
only rituximab
Rituximab or GA101
GA101 is More Effective than Rituximab in Depleting B Cells in Clonus Monkeys
G R
Of Trumps Rituximab in a Clinical Trial for Chronic lymphocytic Leukemia
One Last Look at Where We would be Without Therapeutic Antibodies
Imatinib and Beyond Protein Kinase Inhibitors as Cancer Therapeutics
Protein Kinases as Drug Targets
Problem
There are over 500 protein kinases encoded by the human genome Can a drug target just one
Cancer Drug Gleevec (Imatinib) Wins Lasker AwardPosted at 343 pm CT on September 14 2009
The shortened Philadelphia chromosome seen in certain leukemias (picture from naturecom)The big science news of the day was the announcement of the Lasker Awards informally thought of as the American version of the Nobel Prize for physiology and medicine This yearrsquos clinical medical research award went to a trio of researchers from Oregon Health amp Science University Sloan-Kettering Cancer Center and drug company Novartis but you could just as easily say it was awarded to a drug the cancer treatment Gleevec (imatinib)
Noyes
sufficient specificity for clinical benefit
Imatinib and Related Drugs Target Philadelphia Chromosome Positive CML
bull BCR-ABL fusion creates a dys-regulated protein kinase that drives the proliferation of CML clones
protein kinase active site
bind ATP
bind target protein substrate
imatinib
specific kinase inhibition
A priori targeting the protein substrate binding site might be expected to provide more specificity buthelliphelliphelliphelliphelliphelliphellip
The Imatinib Experience
bull 82 of patients achieve a complete cytogenetic response (65-85)bull majority achieve a major molecular response (40-60)bull 7 year overall survival rate 90 bull event free survival rate 81 (65-85)
Positives
results from the International Randomized Study of Interferon Versus STI571 (IRIS) trial 7-year follow up
Blood (2008) 112 186
various studies from 2003-2008
bull Imatinib however does not destroy the cancer stem cell clone
bull If you remove imatinib CML returns
bull Imatinib is the perfect drug because once you are on it you are on it forever
If you place a cancer cell under selective pressure for long enough without killing it what will happen
- Slide 1
- Slide 2
- Slide 3
- Slide 4
- Slide 5
- Slide 6
- Slide 7
- Slide 8
- Slide 9
- Slide 10
- Slide 11
- Slide 12
- Slide 13
- Slide 14
- Slide 15
- Slide 16
- Slide 17
- Slide 18
- Slide 19
- Slide 20
- Slide 21
- Slide 22
- Slide 23
- Slide 24
-
Interaction of Immunoglobulins with FcγRIII Receptors
polymorphic position 158
AgAg
bivalent
Cold turkey ndash describes the actions of a person who abruptly gives up a habit or addiction rather than gradually easing the process
Immunoglobulins are Subject to N-Linked Glycosylation
Glycoproteins
N-linked glycosylation ndash oligosaccharides linked to Asn residues in the sequence context
Asn-X-Ser or Asn-X-Thr
Proteins are initially modified by a 14 residues core oligosaccharide
Dolichol is a lipid
CH2
C ONSugar
N-glycosidic bond
Hunusual high energy phosphate
ester linkage provides energy for forming N-glycosidic bond
Cotranslational Addition of the Core Oligosaccharide
oligosaccharide transferase
signal peptide
cytoplasm
rough ER membrane
ER lumen
complex
high mannose
Asn-X-SerAsn-X-Thr
Processing the Core Oligosaccharide
Glucose
Mannose
N-acetyl glucosamine
Fucose
Galactose
Sialic acid
High mannose class switches from trimming to addition
Complex class switches from trimming to addition
Antibodies are Glycoproteins and Glycosylation can Influence Effector Function
Antibodies are subject to N-linked glycosylation at residue 297 of the Fc region
Modifying Glycosylation Patterns
Biantennary fucosylated complex oligosaccharide
bull 111308914-N-acetylglucosaminyltransferase III (GnT-III)
GluNAc
Man
Fuc
bull 1113089Man-2
Transfect cells with
After glyco-engineering bisected non-fucosylated antibody
Kinetic Constants for the Binding of Antibodies with Modified Carbohydrates to FcγRIIIa
20 of patients are homozygous Valine
35 of patients are homozygous Phenyalanine
Biacor Traditional equilibrium
binding assay
bull GA101 with native glycosylation pattern binds much better to polymorphic variant with Val
bull GA101 with modified glycosylation pattern binds to either polymorphic variant with roughly equal affinity
Cell Death Assays Annexin Staining
Phosphatidylserine (PS) normally found on cytoplasmic surface of plasma membrane
During intermediate stages of apoptosis PS is translocated to the outer surface of the plasma membrane
Externalization of PS makes it available for binding to fluorescent PS-binding proteins (Annexins)
Added one additional mutation (V for L) in the hinge region
Selected on the basis of ADCC rather than affinity
GA101 Does Not Cause CD20 to Aggregate in Lipid Rafts Like Rituximab
Antibodies to CD20 fall into two classes
bull Class 1 ndash includes Rituximab causes CD20 to aggregate in lipid rafts
bull Class 2 ndash no aggregation fewer molecules bound per cell
G R
Rituximab is more Effective than GA101 at Promoting Complement-Dependent Cell Lysis
rituximab
GA101
GA101 is more Effective at ADCC and B Cell Depletion than Rituximab
GA101
rituximab
GA101 is more Effective than Rituximab in Mouse Xenograph Models of Diffuse B Cell Lymphoma
only rituximab
Rituximab or GA101
GA101 is More Effective than Rituximab in Depleting B Cells in Clonus Monkeys
G R
Of Trumps Rituximab in a Clinical Trial for Chronic lymphocytic Leukemia
One Last Look at Where We would be Without Therapeutic Antibodies
Imatinib and Beyond Protein Kinase Inhibitors as Cancer Therapeutics
Protein Kinases as Drug Targets
Problem
There are over 500 protein kinases encoded by the human genome Can a drug target just one
Cancer Drug Gleevec (Imatinib) Wins Lasker AwardPosted at 343 pm CT on September 14 2009
The shortened Philadelphia chromosome seen in certain leukemias (picture from naturecom)The big science news of the day was the announcement of the Lasker Awards informally thought of as the American version of the Nobel Prize for physiology and medicine This yearrsquos clinical medical research award went to a trio of researchers from Oregon Health amp Science University Sloan-Kettering Cancer Center and drug company Novartis but you could just as easily say it was awarded to a drug the cancer treatment Gleevec (imatinib)
Noyes
sufficient specificity for clinical benefit
Imatinib and Related Drugs Target Philadelphia Chromosome Positive CML
bull BCR-ABL fusion creates a dys-regulated protein kinase that drives the proliferation of CML clones
protein kinase active site
bind ATP
bind target protein substrate
imatinib
specific kinase inhibition
A priori targeting the protein substrate binding site might be expected to provide more specificity buthelliphelliphelliphelliphelliphelliphellip
The Imatinib Experience
bull 82 of patients achieve a complete cytogenetic response (65-85)bull majority achieve a major molecular response (40-60)bull 7 year overall survival rate 90 bull event free survival rate 81 (65-85)
Positives
results from the International Randomized Study of Interferon Versus STI571 (IRIS) trial 7-year follow up
Blood (2008) 112 186
various studies from 2003-2008
bull Imatinib however does not destroy the cancer stem cell clone
bull If you remove imatinib CML returns
bull Imatinib is the perfect drug because once you are on it you are on it forever
If you place a cancer cell under selective pressure for long enough without killing it what will happen
- Slide 1
- Slide 2
- Slide 3
- Slide 4
- Slide 5
- Slide 6
- Slide 7
- Slide 8
- Slide 9
- Slide 10
- Slide 11
- Slide 12
- Slide 13
- Slide 14
- Slide 15
- Slide 16
- Slide 17
- Slide 18
- Slide 19
- Slide 20
- Slide 21
- Slide 22
- Slide 23
- Slide 24
-
Cold turkey ndash describes the actions of a person who abruptly gives up a habit or addiction rather than gradually easing the process
Immunoglobulins are Subject to N-Linked Glycosylation
Glycoproteins
N-linked glycosylation ndash oligosaccharides linked to Asn residues in the sequence context
Asn-X-Ser or Asn-X-Thr
Proteins are initially modified by a 14 residues core oligosaccharide
Dolichol is a lipid
CH2
C ONSugar
N-glycosidic bond
Hunusual high energy phosphate
ester linkage provides energy for forming N-glycosidic bond
Cotranslational Addition of the Core Oligosaccharide
oligosaccharide transferase
signal peptide
cytoplasm
rough ER membrane
ER lumen
complex
high mannose
Asn-X-SerAsn-X-Thr
Processing the Core Oligosaccharide
Glucose
Mannose
N-acetyl glucosamine
Fucose
Galactose
Sialic acid
High mannose class switches from trimming to addition
Complex class switches from trimming to addition
Antibodies are Glycoproteins and Glycosylation can Influence Effector Function
Antibodies are subject to N-linked glycosylation at residue 297 of the Fc region
Modifying Glycosylation Patterns
Biantennary fucosylated complex oligosaccharide
bull 111308914-N-acetylglucosaminyltransferase III (GnT-III)
GluNAc
Man
Fuc
bull 1113089Man-2
Transfect cells with
After glyco-engineering bisected non-fucosylated antibody
Kinetic Constants for the Binding of Antibodies with Modified Carbohydrates to FcγRIIIa
20 of patients are homozygous Valine
35 of patients are homozygous Phenyalanine
Biacor Traditional equilibrium
binding assay
bull GA101 with native glycosylation pattern binds much better to polymorphic variant with Val
bull GA101 with modified glycosylation pattern binds to either polymorphic variant with roughly equal affinity
Cell Death Assays Annexin Staining
Phosphatidylserine (PS) normally found on cytoplasmic surface of plasma membrane
During intermediate stages of apoptosis PS is translocated to the outer surface of the plasma membrane
Externalization of PS makes it available for binding to fluorescent PS-binding proteins (Annexins)
Added one additional mutation (V for L) in the hinge region
Selected on the basis of ADCC rather than affinity
GA101 Does Not Cause CD20 to Aggregate in Lipid Rafts Like Rituximab
Antibodies to CD20 fall into two classes
bull Class 1 ndash includes Rituximab causes CD20 to aggregate in lipid rafts
bull Class 2 ndash no aggregation fewer molecules bound per cell
G R
Rituximab is more Effective than GA101 at Promoting Complement-Dependent Cell Lysis
rituximab
GA101
GA101 is more Effective at ADCC and B Cell Depletion than Rituximab
GA101
rituximab
GA101 is more Effective than Rituximab in Mouse Xenograph Models of Diffuse B Cell Lymphoma
only rituximab
Rituximab or GA101
GA101 is More Effective than Rituximab in Depleting B Cells in Clonus Monkeys
G R
Of Trumps Rituximab in a Clinical Trial for Chronic lymphocytic Leukemia
One Last Look at Where We would be Without Therapeutic Antibodies
Imatinib and Beyond Protein Kinase Inhibitors as Cancer Therapeutics
Protein Kinases as Drug Targets
Problem
There are over 500 protein kinases encoded by the human genome Can a drug target just one
Cancer Drug Gleevec (Imatinib) Wins Lasker AwardPosted at 343 pm CT on September 14 2009
The shortened Philadelphia chromosome seen in certain leukemias (picture from naturecom)The big science news of the day was the announcement of the Lasker Awards informally thought of as the American version of the Nobel Prize for physiology and medicine This yearrsquos clinical medical research award went to a trio of researchers from Oregon Health amp Science University Sloan-Kettering Cancer Center and drug company Novartis but you could just as easily say it was awarded to a drug the cancer treatment Gleevec (imatinib)
Noyes
sufficient specificity for clinical benefit
Imatinib and Related Drugs Target Philadelphia Chromosome Positive CML
bull BCR-ABL fusion creates a dys-regulated protein kinase that drives the proliferation of CML clones
protein kinase active site
bind ATP
bind target protein substrate
imatinib
specific kinase inhibition
A priori targeting the protein substrate binding site might be expected to provide more specificity buthelliphelliphelliphelliphelliphelliphellip
The Imatinib Experience
bull 82 of patients achieve a complete cytogenetic response (65-85)bull majority achieve a major molecular response (40-60)bull 7 year overall survival rate 90 bull event free survival rate 81 (65-85)
Positives
results from the International Randomized Study of Interferon Versus STI571 (IRIS) trial 7-year follow up
Blood (2008) 112 186
various studies from 2003-2008
bull Imatinib however does not destroy the cancer stem cell clone
bull If you remove imatinib CML returns
bull Imatinib is the perfect drug because once you are on it you are on it forever
If you place a cancer cell under selective pressure for long enough without killing it what will happen
- Slide 1
- Slide 2
- Slide 3
- Slide 4
- Slide 5
- Slide 6
- Slide 7
- Slide 8
- Slide 9
- Slide 10
- Slide 11
- Slide 12
- Slide 13
- Slide 14
- Slide 15
- Slide 16
- Slide 17
- Slide 18
- Slide 19
- Slide 20
- Slide 21
- Slide 22
- Slide 23
- Slide 24
-
Glycoproteins
N-linked glycosylation ndash oligosaccharides linked to Asn residues in the sequence context
Asn-X-Ser or Asn-X-Thr
Proteins are initially modified by a 14 residues core oligosaccharide
Dolichol is a lipid
CH2
C ONSugar
N-glycosidic bond
Hunusual high energy phosphate
ester linkage provides energy for forming N-glycosidic bond
Cotranslational Addition of the Core Oligosaccharide
oligosaccharide transferase
signal peptide
cytoplasm
rough ER membrane
ER lumen
complex
high mannose
Asn-X-SerAsn-X-Thr
Processing the Core Oligosaccharide
Glucose
Mannose
N-acetyl glucosamine
Fucose
Galactose
Sialic acid
High mannose class switches from trimming to addition
Complex class switches from trimming to addition
Antibodies are Glycoproteins and Glycosylation can Influence Effector Function
Antibodies are subject to N-linked glycosylation at residue 297 of the Fc region
Modifying Glycosylation Patterns
Biantennary fucosylated complex oligosaccharide
bull 111308914-N-acetylglucosaminyltransferase III (GnT-III)
GluNAc
Man
Fuc
bull 1113089Man-2
Transfect cells with
After glyco-engineering bisected non-fucosylated antibody
Kinetic Constants for the Binding of Antibodies with Modified Carbohydrates to FcγRIIIa
20 of patients are homozygous Valine
35 of patients are homozygous Phenyalanine
Biacor Traditional equilibrium
binding assay
bull GA101 with native glycosylation pattern binds much better to polymorphic variant with Val
bull GA101 with modified glycosylation pattern binds to either polymorphic variant with roughly equal affinity
Cell Death Assays Annexin Staining
Phosphatidylserine (PS) normally found on cytoplasmic surface of plasma membrane
During intermediate stages of apoptosis PS is translocated to the outer surface of the plasma membrane
Externalization of PS makes it available for binding to fluorescent PS-binding proteins (Annexins)
Added one additional mutation (V for L) in the hinge region
Selected on the basis of ADCC rather than affinity
GA101 Does Not Cause CD20 to Aggregate in Lipid Rafts Like Rituximab
Antibodies to CD20 fall into two classes
bull Class 1 ndash includes Rituximab causes CD20 to aggregate in lipid rafts
bull Class 2 ndash no aggregation fewer molecules bound per cell
G R
Rituximab is more Effective than GA101 at Promoting Complement-Dependent Cell Lysis
rituximab
GA101
GA101 is more Effective at ADCC and B Cell Depletion than Rituximab
GA101
rituximab
GA101 is more Effective than Rituximab in Mouse Xenograph Models of Diffuse B Cell Lymphoma
only rituximab
Rituximab or GA101
GA101 is More Effective than Rituximab in Depleting B Cells in Clonus Monkeys
G R
Of Trumps Rituximab in a Clinical Trial for Chronic lymphocytic Leukemia
One Last Look at Where We would be Without Therapeutic Antibodies
Imatinib and Beyond Protein Kinase Inhibitors as Cancer Therapeutics
Protein Kinases as Drug Targets
Problem
There are over 500 protein kinases encoded by the human genome Can a drug target just one
Cancer Drug Gleevec (Imatinib) Wins Lasker AwardPosted at 343 pm CT on September 14 2009
The shortened Philadelphia chromosome seen in certain leukemias (picture from naturecom)The big science news of the day was the announcement of the Lasker Awards informally thought of as the American version of the Nobel Prize for physiology and medicine This yearrsquos clinical medical research award went to a trio of researchers from Oregon Health amp Science University Sloan-Kettering Cancer Center and drug company Novartis but you could just as easily say it was awarded to a drug the cancer treatment Gleevec (imatinib)
Noyes
sufficient specificity for clinical benefit
Imatinib and Related Drugs Target Philadelphia Chromosome Positive CML
bull BCR-ABL fusion creates a dys-regulated protein kinase that drives the proliferation of CML clones
protein kinase active site
bind ATP
bind target protein substrate
imatinib
specific kinase inhibition
A priori targeting the protein substrate binding site might be expected to provide more specificity buthelliphelliphelliphelliphelliphelliphellip
The Imatinib Experience
bull 82 of patients achieve a complete cytogenetic response (65-85)bull majority achieve a major molecular response (40-60)bull 7 year overall survival rate 90 bull event free survival rate 81 (65-85)
Positives
results from the International Randomized Study of Interferon Versus STI571 (IRIS) trial 7-year follow up
Blood (2008) 112 186
various studies from 2003-2008
bull Imatinib however does not destroy the cancer stem cell clone
bull If you remove imatinib CML returns
bull Imatinib is the perfect drug because once you are on it you are on it forever
If you place a cancer cell under selective pressure for long enough without killing it what will happen
- Slide 1
- Slide 2
- Slide 3
- Slide 4
- Slide 5
- Slide 6
- Slide 7
- Slide 8
- Slide 9
- Slide 10
- Slide 11
- Slide 12
- Slide 13
- Slide 14
- Slide 15
- Slide 16
- Slide 17
- Slide 18
- Slide 19
- Slide 20
- Slide 21
- Slide 22
- Slide 23
- Slide 24
-
Cotranslational Addition of the Core Oligosaccharide
oligosaccharide transferase
signal peptide
cytoplasm
rough ER membrane
ER lumen
complex
high mannose
Asn-X-SerAsn-X-Thr
Processing the Core Oligosaccharide
Glucose
Mannose
N-acetyl glucosamine
Fucose
Galactose
Sialic acid
High mannose class switches from trimming to addition
Complex class switches from trimming to addition
Antibodies are Glycoproteins and Glycosylation can Influence Effector Function
Antibodies are subject to N-linked glycosylation at residue 297 of the Fc region
Modifying Glycosylation Patterns
Biantennary fucosylated complex oligosaccharide
bull 111308914-N-acetylglucosaminyltransferase III (GnT-III)
GluNAc
Man
Fuc
bull 1113089Man-2
Transfect cells with
After glyco-engineering bisected non-fucosylated antibody
Kinetic Constants for the Binding of Antibodies with Modified Carbohydrates to FcγRIIIa
20 of patients are homozygous Valine
35 of patients are homozygous Phenyalanine
Biacor Traditional equilibrium
binding assay
bull GA101 with native glycosylation pattern binds much better to polymorphic variant with Val
bull GA101 with modified glycosylation pattern binds to either polymorphic variant with roughly equal affinity
Cell Death Assays Annexin Staining
Phosphatidylserine (PS) normally found on cytoplasmic surface of plasma membrane
During intermediate stages of apoptosis PS is translocated to the outer surface of the plasma membrane
Externalization of PS makes it available for binding to fluorescent PS-binding proteins (Annexins)
Added one additional mutation (V for L) in the hinge region
Selected on the basis of ADCC rather than affinity
GA101 Does Not Cause CD20 to Aggregate in Lipid Rafts Like Rituximab
Antibodies to CD20 fall into two classes
bull Class 1 ndash includes Rituximab causes CD20 to aggregate in lipid rafts
bull Class 2 ndash no aggregation fewer molecules bound per cell
G R
Rituximab is more Effective than GA101 at Promoting Complement-Dependent Cell Lysis
rituximab
GA101
GA101 is more Effective at ADCC and B Cell Depletion than Rituximab
GA101
rituximab
GA101 is more Effective than Rituximab in Mouse Xenograph Models of Diffuse B Cell Lymphoma
only rituximab
Rituximab or GA101
GA101 is More Effective than Rituximab in Depleting B Cells in Clonus Monkeys
G R
Of Trumps Rituximab in a Clinical Trial for Chronic lymphocytic Leukemia
One Last Look at Where We would be Without Therapeutic Antibodies
Imatinib and Beyond Protein Kinase Inhibitors as Cancer Therapeutics
Protein Kinases as Drug Targets
Problem
There are over 500 protein kinases encoded by the human genome Can a drug target just one
Cancer Drug Gleevec (Imatinib) Wins Lasker AwardPosted at 343 pm CT on September 14 2009
The shortened Philadelphia chromosome seen in certain leukemias (picture from naturecom)The big science news of the day was the announcement of the Lasker Awards informally thought of as the American version of the Nobel Prize for physiology and medicine This yearrsquos clinical medical research award went to a trio of researchers from Oregon Health amp Science University Sloan-Kettering Cancer Center and drug company Novartis but you could just as easily say it was awarded to a drug the cancer treatment Gleevec (imatinib)
Noyes
sufficient specificity for clinical benefit
Imatinib and Related Drugs Target Philadelphia Chromosome Positive CML
bull BCR-ABL fusion creates a dys-regulated protein kinase that drives the proliferation of CML clones
protein kinase active site
bind ATP
bind target protein substrate
imatinib
specific kinase inhibition
A priori targeting the protein substrate binding site might be expected to provide more specificity buthelliphelliphelliphelliphelliphelliphellip
The Imatinib Experience
bull 82 of patients achieve a complete cytogenetic response (65-85)bull majority achieve a major molecular response (40-60)bull 7 year overall survival rate 90 bull event free survival rate 81 (65-85)
Positives
results from the International Randomized Study of Interferon Versus STI571 (IRIS) trial 7-year follow up
Blood (2008) 112 186
various studies from 2003-2008
bull Imatinib however does not destroy the cancer stem cell clone
bull If you remove imatinib CML returns
bull Imatinib is the perfect drug because once you are on it you are on it forever
If you place a cancer cell under selective pressure for long enough without killing it what will happen
- Slide 1
- Slide 2
- Slide 3
- Slide 4
- Slide 5
- Slide 6
- Slide 7
- Slide 8
- Slide 9
- Slide 10
- Slide 11
- Slide 12
- Slide 13
- Slide 14
- Slide 15
- Slide 16
- Slide 17
- Slide 18
- Slide 19
- Slide 20
- Slide 21
- Slide 22
- Slide 23
- Slide 24
-
Processing the Core Oligosaccharide
Glucose
Mannose
N-acetyl glucosamine
Fucose
Galactose
Sialic acid
High mannose class switches from trimming to addition
Complex class switches from trimming to addition
Antibodies are Glycoproteins and Glycosylation can Influence Effector Function
Antibodies are subject to N-linked glycosylation at residue 297 of the Fc region
Modifying Glycosylation Patterns
Biantennary fucosylated complex oligosaccharide
bull 111308914-N-acetylglucosaminyltransferase III (GnT-III)
GluNAc
Man
Fuc
bull 1113089Man-2
Transfect cells with
After glyco-engineering bisected non-fucosylated antibody
Kinetic Constants for the Binding of Antibodies with Modified Carbohydrates to FcγRIIIa
20 of patients are homozygous Valine
35 of patients are homozygous Phenyalanine
Biacor Traditional equilibrium
binding assay
bull GA101 with native glycosylation pattern binds much better to polymorphic variant with Val
bull GA101 with modified glycosylation pattern binds to either polymorphic variant with roughly equal affinity
Cell Death Assays Annexin Staining
Phosphatidylserine (PS) normally found on cytoplasmic surface of plasma membrane
During intermediate stages of apoptosis PS is translocated to the outer surface of the plasma membrane
Externalization of PS makes it available for binding to fluorescent PS-binding proteins (Annexins)
Added one additional mutation (V for L) in the hinge region
Selected on the basis of ADCC rather than affinity
GA101 Does Not Cause CD20 to Aggregate in Lipid Rafts Like Rituximab
Antibodies to CD20 fall into two classes
bull Class 1 ndash includes Rituximab causes CD20 to aggregate in lipid rafts
bull Class 2 ndash no aggregation fewer molecules bound per cell
G R
Rituximab is more Effective than GA101 at Promoting Complement-Dependent Cell Lysis
rituximab
GA101
GA101 is more Effective at ADCC and B Cell Depletion than Rituximab
GA101
rituximab
GA101 is more Effective than Rituximab in Mouse Xenograph Models of Diffuse B Cell Lymphoma
only rituximab
Rituximab or GA101
GA101 is More Effective than Rituximab in Depleting B Cells in Clonus Monkeys
G R
Of Trumps Rituximab in a Clinical Trial for Chronic lymphocytic Leukemia
One Last Look at Where We would be Without Therapeutic Antibodies
Imatinib and Beyond Protein Kinase Inhibitors as Cancer Therapeutics
Protein Kinases as Drug Targets
Problem
There are over 500 protein kinases encoded by the human genome Can a drug target just one
Cancer Drug Gleevec (Imatinib) Wins Lasker AwardPosted at 343 pm CT on September 14 2009
The shortened Philadelphia chromosome seen in certain leukemias (picture from naturecom)The big science news of the day was the announcement of the Lasker Awards informally thought of as the American version of the Nobel Prize for physiology and medicine This yearrsquos clinical medical research award went to a trio of researchers from Oregon Health amp Science University Sloan-Kettering Cancer Center and drug company Novartis but you could just as easily say it was awarded to a drug the cancer treatment Gleevec (imatinib)
Noyes
sufficient specificity for clinical benefit
Imatinib and Related Drugs Target Philadelphia Chromosome Positive CML
bull BCR-ABL fusion creates a dys-regulated protein kinase that drives the proliferation of CML clones
protein kinase active site
bind ATP
bind target protein substrate
imatinib
specific kinase inhibition
A priori targeting the protein substrate binding site might be expected to provide more specificity buthelliphelliphelliphelliphelliphelliphellip
The Imatinib Experience
bull 82 of patients achieve a complete cytogenetic response (65-85)bull majority achieve a major molecular response (40-60)bull 7 year overall survival rate 90 bull event free survival rate 81 (65-85)
Positives
results from the International Randomized Study of Interferon Versus STI571 (IRIS) trial 7-year follow up
Blood (2008) 112 186
various studies from 2003-2008
bull Imatinib however does not destroy the cancer stem cell clone
bull If you remove imatinib CML returns
bull Imatinib is the perfect drug because once you are on it you are on it forever
If you place a cancer cell under selective pressure for long enough without killing it what will happen
- Slide 1
- Slide 2
- Slide 3
- Slide 4
- Slide 5
- Slide 6
- Slide 7
- Slide 8
- Slide 9
- Slide 10
- Slide 11
- Slide 12
- Slide 13
- Slide 14
- Slide 15
- Slide 16
- Slide 17
- Slide 18
- Slide 19
- Slide 20
- Slide 21
- Slide 22
- Slide 23
- Slide 24
-
Antibodies are Glycoproteins and Glycosylation can Influence Effector Function
Antibodies are subject to N-linked glycosylation at residue 297 of the Fc region
Modifying Glycosylation Patterns
Biantennary fucosylated complex oligosaccharide
bull 111308914-N-acetylglucosaminyltransferase III (GnT-III)
GluNAc
Man
Fuc
bull 1113089Man-2
Transfect cells with
After glyco-engineering bisected non-fucosylated antibody
Kinetic Constants for the Binding of Antibodies with Modified Carbohydrates to FcγRIIIa
20 of patients are homozygous Valine
35 of patients are homozygous Phenyalanine
Biacor Traditional equilibrium
binding assay
bull GA101 with native glycosylation pattern binds much better to polymorphic variant with Val
bull GA101 with modified glycosylation pattern binds to either polymorphic variant with roughly equal affinity
Cell Death Assays Annexin Staining
Phosphatidylserine (PS) normally found on cytoplasmic surface of plasma membrane
During intermediate stages of apoptosis PS is translocated to the outer surface of the plasma membrane
Externalization of PS makes it available for binding to fluorescent PS-binding proteins (Annexins)
Added one additional mutation (V for L) in the hinge region
Selected on the basis of ADCC rather than affinity
GA101 Does Not Cause CD20 to Aggregate in Lipid Rafts Like Rituximab
Antibodies to CD20 fall into two classes
bull Class 1 ndash includes Rituximab causes CD20 to aggregate in lipid rafts
bull Class 2 ndash no aggregation fewer molecules bound per cell
G R
Rituximab is more Effective than GA101 at Promoting Complement-Dependent Cell Lysis
rituximab
GA101
GA101 is more Effective at ADCC and B Cell Depletion than Rituximab
GA101
rituximab
GA101 is more Effective than Rituximab in Mouse Xenograph Models of Diffuse B Cell Lymphoma
only rituximab
Rituximab or GA101
GA101 is More Effective than Rituximab in Depleting B Cells in Clonus Monkeys
G R
Of Trumps Rituximab in a Clinical Trial for Chronic lymphocytic Leukemia
One Last Look at Where We would be Without Therapeutic Antibodies
Imatinib and Beyond Protein Kinase Inhibitors as Cancer Therapeutics
Protein Kinases as Drug Targets
Problem
There are over 500 protein kinases encoded by the human genome Can a drug target just one
Cancer Drug Gleevec (Imatinib) Wins Lasker AwardPosted at 343 pm CT on September 14 2009
The shortened Philadelphia chromosome seen in certain leukemias (picture from naturecom)The big science news of the day was the announcement of the Lasker Awards informally thought of as the American version of the Nobel Prize for physiology and medicine This yearrsquos clinical medical research award went to a trio of researchers from Oregon Health amp Science University Sloan-Kettering Cancer Center and drug company Novartis but you could just as easily say it was awarded to a drug the cancer treatment Gleevec (imatinib)
Noyes
sufficient specificity for clinical benefit
Imatinib and Related Drugs Target Philadelphia Chromosome Positive CML
bull BCR-ABL fusion creates a dys-regulated protein kinase that drives the proliferation of CML clones
protein kinase active site
bind ATP
bind target protein substrate
imatinib
specific kinase inhibition
A priori targeting the protein substrate binding site might be expected to provide more specificity buthelliphelliphelliphelliphelliphelliphellip
The Imatinib Experience
bull 82 of patients achieve a complete cytogenetic response (65-85)bull majority achieve a major molecular response (40-60)bull 7 year overall survival rate 90 bull event free survival rate 81 (65-85)
Positives
results from the International Randomized Study of Interferon Versus STI571 (IRIS) trial 7-year follow up
Blood (2008) 112 186
various studies from 2003-2008
bull Imatinib however does not destroy the cancer stem cell clone
bull If you remove imatinib CML returns
bull Imatinib is the perfect drug because once you are on it you are on it forever
If you place a cancer cell under selective pressure for long enough without killing it what will happen
- Slide 1
- Slide 2
- Slide 3
- Slide 4
- Slide 5
- Slide 6
- Slide 7
- Slide 8
- Slide 9
- Slide 10
- Slide 11
- Slide 12
- Slide 13
- Slide 14
- Slide 15
- Slide 16
- Slide 17
- Slide 18
- Slide 19
- Slide 20
- Slide 21
- Slide 22
- Slide 23
- Slide 24
-
Modifying Glycosylation Patterns
Biantennary fucosylated complex oligosaccharide
bull 111308914-N-acetylglucosaminyltransferase III (GnT-III)
GluNAc
Man
Fuc
bull 1113089Man-2
Transfect cells with
After glyco-engineering bisected non-fucosylated antibody
Kinetic Constants for the Binding of Antibodies with Modified Carbohydrates to FcγRIIIa
20 of patients are homozygous Valine
35 of patients are homozygous Phenyalanine
Biacor Traditional equilibrium
binding assay
bull GA101 with native glycosylation pattern binds much better to polymorphic variant with Val
bull GA101 with modified glycosylation pattern binds to either polymorphic variant with roughly equal affinity
Cell Death Assays Annexin Staining
Phosphatidylserine (PS) normally found on cytoplasmic surface of plasma membrane
During intermediate stages of apoptosis PS is translocated to the outer surface of the plasma membrane
Externalization of PS makes it available for binding to fluorescent PS-binding proteins (Annexins)
Added one additional mutation (V for L) in the hinge region
Selected on the basis of ADCC rather than affinity
GA101 Does Not Cause CD20 to Aggregate in Lipid Rafts Like Rituximab
Antibodies to CD20 fall into two classes
bull Class 1 ndash includes Rituximab causes CD20 to aggregate in lipid rafts
bull Class 2 ndash no aggregation fewer molecules bound per cell
G R
Rituximab is more Effective than GA101 at Promoting Complement-Dependent Cell Lysis
rituximab
GA101
GA101 is more Effective at ADCC and B Cell Depletion than Rituximab
GA101
rituximab
GA101 is more Effective than Rituximab in Mouse Xenograph Models of Diffuse B Cell Lymphoma
only rituximab
Rituximab or GA101
GA101 is More Effective than Rituximab in Depleting B Cells in Clonus Monkeys
G R
Of Trumps Rituximab in a Clinical Trial for Chronic lymphocytic Leukemia
One Last Look at Where We would be Without Therapeutic Antibodies
Imatinib and Beyond Protein Kinase Inhibitors as Cancer Therapeutics
Protein Kinases as Drug Targets
Problem
There are over 500 protein kinases encoded by the human genome Can a drug target just one
Cancer Drug Gleevec (Imatinib) Wins Lasker AwardPosted at 343 pm CT on September 14 2009
The shortened Philadelphia chromosome seen in certain leukemias (picture from naturecom)The big science news of the day was the announcement of the Lasker Awards informally thought of as the American version of the Nobel Prize for physiology and medicine This yearrsquos clinical medical research award went to a trio of researchers from Oregon Health amp Science University Sloan-Kettering Cancer Center and drug company Novartis but you could just as easily say it was awarded to a drug the cancer treatment Gleevec (imatinib)
Noyes
sufficient specificity for clinical benefit
Imatinib and Related Drugs Target Philadelphia Chromosome Positive CML
bull BCR-ABL fusion creates a dys-regulated protein kinase that drives the proliferation of CML clones
protein kinase active site
bind ATP
bind target protein substrate
imatinib
specific kinase inhibition
A priori targeting the protein substrate binding site might be expected to provide more specificity buthelliphelliphelliphelliphelliphelliphellip
The Imatinib Experience
bull 82 of patients achieve a complete cytogenetic response (65-85)bull majority achieve a major molecular response (40-60)bull 7 year overall survival rate 90 bull event free survival rate 81 (65-85)
Positives
results from the International Randomized Study of Interferon Versus STI571 (IRIS) trial 7-year follow up
Blood (2008) 112 186
various studies from 2003-2008
bull Imatinib however does not destroy the cancer stem cell clone
bull If you remove imatinib CML returns
bull Imatinib is the perfect drug because once you are on it you are on it forever
If you place a cancer cell under selective pressure for long enough without killing it what will happen
- Slide 1
- Slide 2
- Slide 3
- Slide 4
- Slide 5
- Slide 6
- Slide 7
- Slide 8
- Slide 9
- Slide 10
- Slide 11
- Slide 12
- Slide 13
- Slide 14
- Slide 15
- Slide 16
- Slide 17
- Slide 18
- Slide 19
- Slide 20
- Slide 21
- Slide 22
- Slide 23
- Slide 24
-
Kinetic Constants for the Binding of Antibodies with Modified Carbohydrates to FcγRIIIa
20 of patients are homozygous Valine
35 of patients are homozygous Phenyalanine
Biacor Traditional equilibrium
binding assay
bull GA101 with native glycosylation pattern binds much better to polymorphic variant with Val
bull GA101 with modified glycosylation pattern binds to either polymorphic variant with roughly equal affinity
Cell Death Assays Annexin Staining
Phosphatidylserine (PS) normally found on cytoplasmic surface of plasma membrane
During intermediate stages of apoptosis PS is translocated to the outer surface of the plasma membrane
Externalization of PS makes it available for binding to fluorescent PS-binding proteins (Annexins)
Added one additional mutation (V for L) in the hinge region
Selected on the basis of ADCC rather than affinity
GA101 Does Not Cause CD20 to Aggregate in Lipid Rafts Like Rituximab
Antibodies to CD20 fall into two classes
bull Class 1 ndash includes Rituximab causes CD20 to aggregate in lipid rafts
bull Class 2 ndash no aggregation fewer molecules bound per cell
G R
Rituximab is more Effective than GA101 at Promoting Complement-Dependent Cell Lysis
rituximab
GA101
GA101 is more Effective at ADCC and B Cell Depletion than Rituximab
GA101
rituximab
GA101 is more Effective than Rituximab in Mouse Xenograph Models of Diffuse B Cell Lymphoma
only rituximab
Rituximab or GA101
GA101 is More Effective than Rituximab in Depleting B Cells in Clonus Monkeys
G R
Of Trumps Rituximab in a Clinical Trial for Chronic lymphocytic Leukemia
One Last Look at Where We would be Without Therapeutic Antibodies
Imatinib and Beyond Protein Kinase Inhibitors as Cancer Therapeutics
Protein Kinases as Drug Targets
Problem
There are over 500 protein kinases encoded by the human genome Can a drug target just one
Cancer Drug Gleevec (Imatinib) Wins Lasker AwardPosted at 343 pm CT on September 14 2009
The shortened Philadelphia chromosome seen in certain leukemias (picture from naturecom)The big science news of the day was the announcement of the Lasker Awards informally thought of as the American version of the Nobel Prize for physiology and medicine This yearrsquos clinical medical research award went to a trio of researchers from Oregon Health amp Science University Sloan-Kettering Cancer Center and drug company Novartis but you could just as easily say it was awarded to a drug the cancer treatment Gleevec (imatinib)
Noyes
sufficient specificity for clinical benefit
Imatinib and Related Drugs Target Philadelphia Chromosome Positive CML
bull BCR-ABL fusion creates a dys-regulated protein kinase that drives the proliferation of CML clones
protein kinase active site
bind ATP
bind target protein substrate
imatinib
specific kinase inhibition
A priori targeting the protein substrate binding site might be expected to provide more specificity buthelliphelliphelliphelliphelliphelliphellip
The Imatinib Experience
bull 82 of patients achieve a complete cytogenetic response (65-85)bull majority achieve a major molecular response (40-60)bull 7 year overall survival rate 90 bull event free survival rate 81 (65-85)
Positives
results from the International Randomized Study of Interferon Versus STI571 (IRIS) trial 7-year follow up
Blood (2008) 112 186
various studies from 2003-2008
bull Imatinib however does not destroy the cancer stem cell clone
bull If you remove imatinib CML returns
bull Imatinib is the perfect drug because once you are on it you are on it forever
If you place a cancer cell under selective pressure for long enough without killing it what will happen
- Slide 1
- Slide 2
- Slide 3
- Slide 4
- Slide 5
- Slide 6
- Slide 7
- Slide 8
- Slide 9
- Slide 10
- Slide 11
- Slide 12
- Slide 13
- Slide 14
- Slide 15
- Slide 16
- Slide 17
- Slide 18
- Slide 19
- Slide 20
- Slide 21
- Slide 22
- Slide 23
- Slide 24
-
Cell Death Assays Annexin Staining
Phosphatidylserine (PS) normally found on cytoplasmic surface of plasma membrane
During intermediate stages of apoptosis PS is translocated to the outer surface of the plasma membrane
Externalization of PS makes it available for binding to fluorescent PS-binding proteins (Annexins)
Added one additional mutation (V for L) in the hinge region
Selected on the basis of ADCC rather than affinity
GA101 Does Not Cause CD20 to Aggregate in Lipid Rafts Like Rituximab
Antibodies to CD20 fall into two classes
bull Class 1 ndash includes Rituximab causes CD20 to aggregate in lipid rafts
bull Class 2 ndash no aggregation fewer molecules bound per cell
G R
Rituximab is more Effective than GA101 at Promoting Complement-Dependent Cell Lysis
rituximab
GA101
GA101 is more Effective at ADCC and B Cell Depletion than Rituximab
GA101
rituximab
GA101 is more Effective than Rituximab in Mouse Xenograph Models of Diffuse B Cell Lymphoma
only rituximab
Rituximab or GA101
GA101 is More Effective than Rituximab in Depleting B Cells in Clonus Monkeys
G R
Of Trumps Rituximab in a Clinical Trial for Chronic lymphocytic Leukemia
One Last Look at Where We would be Without Therapeutic Antibodies
Imatinib and Beyond Protein Kinase Inhibitors as Cancer Therapeutics
Protein Kinases as Drug Targets
Problem
There are over 500 protein kinases encoded by the human genome Can a drug target just one
Cancer Drug Gleevec (Imatinib) Wins Lasker AwardPosted at 343 pm CT on September 14 2009
The shortened Philadelphia chromosome seen in certain leukemias (picture from naturecom)The big science news of the day was the announcement of the Lasker Awards informally thought of as the American version of the Nobel Prize for physiology and medicine This yearrsquos clinical medical research award went to a trio of researchers from Oregon Health amp Science University Sloan-Kettering Cancer Center and drug company Novartis but you could just as easily say it was awarded to a drug the cancer treatment Gleevec (imatinib)
Noyes
sufficient specificity for clinical benefit
Imatinib and Related Drugs Target Philadelphia Chromosome Positive CML
bull BCR-ABL fusion creates a dys-regulated protein kinase that drives the proliferation of CML clones
protein kinase active site
bind ATP
bind target protein substrate
imatinib
specific kinase inhibition
A priori targeting the protein substrate binding site might be expected to provide more specificity buthelliphelliphelliphelliphelliphelliphellip
The Imatinib Experience
bull 82 of patients achieve a complete cytogenetic response (65-85)bull majority achieve a major molecular response (40-60)bull 7 year overall survival rate 90 bull event free survival rate 81 (65-85)
Positives
results from the International Randomized Study of Interferon Versus STI571 (IRIS) trial 7-year follow up
Blood (2008) 112 186
various studies from 2003-2008
bull Imatinib however does not destroy the cancer stem cell clone
bull If you remove imatinib CML returns
bull Imatinib is the perfect drug because once you are on it you are on it forever
If you place a cancer cell under selective pressure for long enough without killing it what will happen
- Slide 1
- Slide 2
- Slide 3
- Slide 4
- Slide 5
- Slide 6
- Slide 7
- Slide 8
- Slide 9
- Slide 10
- Slide 11
- Slide 12
- Slide 13
- Slide 14
- Slide 15
- Slide 16
- Slide 17
- Slide 18
- Slide 19
- Slide 20
- Slide 21
- Slide 22
- Slide 23
- Slide 24
-
GA101 Does Not Cause CD20 to Aggregate in Lipid Rafts Like Rituximab
Antibodies to CD20 fall into two classes
bull Class 1 ndash includes Rituximab causes CD20 to aggregate in lipid rafts
bull Class 2 ndash no aggregation fewer molecules bound per cell
G R
Rituximab is more Effective than GA101 at Promoting Complement-Dependent Cell Lysis
rituximab
GA101
GA101 is more Effective at ADCC and B Cell Depletion than Rituximab
GA101
rituximab
GA101 is more Effective than Rituximab in Mouse Xenograph Models of Diffuse B Cell Lymphoma
only rituximab
Rituximab or GA101
GA101 is More Effective than Rituximab in Depleting B Cells in Clonus Monkeys
G R
Of Trumps Rituximab in a Clinical Trial for Chronic lymphocytic Leukemia
One Last Look at Where We would be Without Therapeutic Antibodies
Imatinib and Beyond Protein Kinase Inhibitors as Cancer Therapeutics
Protein Kinases as Drug Targets
Problem
There are over 500 protein kinases encoded by the human genome Can a drug target just one
Cancer Drug Gleevec (Imatinib) Wins Lasker AwardPosted at 343 pm CT on September 14 2009
The shortened Philadelphia chromosome seen in certain leukemias (picture from naturecom)The big science news of the day was the announcement of the Lasker Awards informally thought of as the American version of the Nobel Prize for physiology and medicine This yearrsquos clinical medical research award went to a trio of researchers from Oregon Health amp Science University Sloan-Kettering Cancer Center and drug company Novartis but you could just as easily say it was awarded to a drug the cancer treatment Gleevec (imatinib)
Noyes
sufficient specificity for clinical benefit
Imatinib and Related Drugs Target Philadelphia Chromosome Positive CML
bull BCR-ABL fusion creates a dys-regulated protein kinase that drives the proliferation of CML clones
protein kinase active site
bind ATP
bind target protein substrate
imatinib
specific kinase inhibition
A priori targeting the protein substrate binding site might be expected to provide more specificity buthelliphelliphelliphelliphelliphelliphellip
The Imatinib Experience
bull 82 of patients achieve a complete cytogenetic response (65-85)bull majority achieve a major molecular response (40-60)bull 7 year overall survival rate 90 bull event free survival rate 81 (65-85)
Positives
results from the International Randomized Study of Interferon Versus STI571 (IRIS) trial 7-year follow up
Blood (2008) 112 186
various studies from 2003-2008
bull Imatinib however does not destroy the cancer stem cell clone
bull If you remove imatinib CML returns
bull Imatinib is the perfect drug because once you are on it you are on it forever
If you place a cancer cell under selective pressure for long enough without killing it what will happen
- Slide 1
- Slide 2
- Slide 3
- Slide 4
- Slide 5
- Slide 6
- Slide 7
- Slide 8
- Slide 9
- Slide 10
- Slide 11
- Slide 12
- Slide 13
- Slide 14
- Slide 15
- Slide 16
- Slide 17
- Slide 18
- Slide 19
- Slide 20
- Slide 21
- Slide 22
- Slide 23
- Slide 24
-
Rituximab is more Effective than GA101 at Promoting Complement-Dependent Cell Lysis
rituximab
GA101
GA101 is more Effective at ADCC and B Cell Depletion than Rituximab
GA101
rituximab
GA101 is more Effective than Rituximab in Mouse Xenograph Models of Diffuse B Cell Lymphoma
only rituximab
Rituximab or GA101
GA101 is More Effective than Rituximab in Depleting B Cells in Clonus Monkeys
G R
Of Trumps Rituximab in a Clinical Trial for Chronic lymphocytic Leukemia
One Last Look at Where We would be Without Therapeutic Antibodies
Imatinib and Beyond Protein Kinase Inhibitors as Cancer Therapeutics
Protein Kinases as Drug Targets
Problem
There are over 500 protein kinases encoded by the human genome Can a drug target just one
Cancer Drug Gleevec (Imatinib) Wins Lasker AwardPosted at 343 pm CT on September 14 2009
The shortened Philadelphia chromosome seen in certain leukemias (picture from naturecom)The big science news of the day was the announcement of the Lasker Awards informally thought of as the American version of the Nobel Prize for physiology and medicine This yearrsquos clinical medical research award went to a trio of researchers from Oregon Health amp Science University Sloan-Kettering Cancer Center and drug company Novartis but you could just as easily say it was awarded to a drug the cancer treatment Gleevec (imatinib)
Noyes
sufficient specificity for clinical benefit
Imatinib and Related Drugs Target Philadelphia Chromosome Positive CML
bull BCR-ABL fusion creates a dys-regulated protein kinase that drives the proliferation of CML clones
protein kinase active site
bind ATP
bind target protein substrate
imatinib
specific kinase inhibition
A priori targeting the protein substrate binding site might be expected to provide more specificity buthelliphelliphelliphelliphelliphelliphellip
The Imatinib Experience
bull 82 of patients achieve a complete cytogenetic response (65-85)bull majority achieve a major molecular response (40-60)bull 7 year overall survival rate 90 bull event free survival rate 81 (65-85)
Positives
results from the International Randomized Study of Interferon Versus STI571 (IRIS) trial 7-year follow up
Blood (2008) 112 186
various studies from 2003-2008
bull Imatinib however does not destroy the cancer stem cell clone
bull If you remove imatinib CML returns
bull Imatinib is the perfect drug because once you are on it you are on it forever
If you place a cancer cell under selective pressure for long enough without killing it what will happen
- Slide 1
- Slide 2
- Slide 3
- Slide 4
- Slide 5
- Slide 6
- Slide 7
- Slide 8
- Slide 9
- Slide 10
- Slide 11
- Slide 12
- Slide 13
- Slide 14
- Slide 15
- Slide 16
- Slide 17
- Slide 18
- Slide 19
- Slide 20
- Slide 21
- Slide 22
- Slide 23
- Slide 24
-
GA101 is more Effective at ADCC and B Cell Depletion than Rituximab
GA101
rituximab
GA101 is more Effective than Rituximab in Mouse Xenograph Models of Diffuse B Cell Lymphoma
only rituximab
Rituximab or GA101
GA101 is More Effective than Rituximab in Depleting B Cells in Clonus Monkeys
G R
Of Trumps Rituximab in a Clinical Trial for Chronic lymphocytic Leukemia
One Last Look at Where We would be Without Therapeutic Antibodies
Imatinib and Beyond Protein Kinase Inhibitors as Cancer Therapeutics
Protein Kinases as Drug Targets
Problem
There are over 500 protein kinases encoded by the human genome Can a drug target just one
Cancer Drug Gleevec (Imatinib) Wins Lasker AwardPosted at 343 pm CT on September 14 2009
The shortened Philadelphia chromosome seen in certain leukemias (picture from naturecom)The big science news of the day was the announcement of the Lasker Awards informally thought of as the American version of the Nobel Prize for physiology and medicine This yearrsquos clinical medical research award went to a trio of researchers from Oregon Health amp Science University Sloan-Kettering Cancer Center and drug company Novartis but you could just as easily say it was awarded to a drug the cancer treatment Gleevec (imatinib)
Noyes
sufficient specificity for clinical benefit
Imatinib and Related Drugs Target Philadelphia Chromosome Positive CML
bull BCR-ABL fusion creates a dys-regulated protein kinase that drives the proliferation of CML clones
protein kinase active site
bind ATP
bind target protein substrate
imatinib
specific kinase inhibition
A priori targeting the protein substrate binding site might be expected to provide more specificity buthelliphelliphelliphelliphelliphelliphellip
The Imatinib Experience
bull 82 of patients achieve a complete cytogenetic response (65-85)bull majority achieve a major molecular response (40-60)bull 7 year overall survival rate 90 bull event free survival rate 81 (65-85)
Positives
results from the International Randomized Study of Interferon Versus STI571 (IRIS) trial 7-year follow up
Blood (2008) 112 186
various studies from 2003-2008
bull Imatinib however does not destroy the cancer stem cell clone
bull If you remove imatinib CML returns
bull Imatinib is the perfect drug because once you are on it you are on it forever
If you place a cancer cell under selective pressure for long enough without killing it what will happen
- Slide 1
- Slide 2
- Slide 3
- Slide 4
- Slide 5
- Slide 6
- Slide 7
- Slide 8
- Slide 9
- Slide 10
- Slide 11
- Slide 12
- Slide 13
- Slide 14
- Slide 15
- Slide 16
- Slide 17
- Slide 18
- Slide 19
- Slide 20
- Slide 21
- Slide 22
- Slide 23
- Slide 24
-
GA101 is more Effective than Rituximab in Mouse Xenograph Models of Diffuse B Cell Lymphoma
only rituximab
Rituximab or GA101
GA101 is More Effective than Rituximab in Depleting B Cells in Clonus Monkeys
G R
Of Trumps Rituximab in a Clinical Trial for Chronic lymphocytic Leukemia
One Last Look at Where We would be Without Therapeutic Antibodies
Imatinib and Beyond Protein Kinase Inhibitors as Cancer Therapeutics
Protein Kinases as Drug Targets
Problem
There are over 500 protein kinases encoded by the human genome Can a drug target just one
Cancer Drug Gleevec (Imatinib) Wins Lasker AwardPosted at 343 pm CT on September 14 2009
The shortened Philadelphia chromosome seen in certain leukemias (picture from naturecom)The big science news of the day was the announcement of the Lasker Awards informally thought of as the American version of the Nobel Prize for physiology and medicine This yearrsquos clinical medical research award went to a trio of researchers from Oregon Health amp Science University Sloan-Kettering Cancer Center and drug company Novartis but you could just as easily say it was awarded to a drug the cancer treatment Gleevec (imatinib)
Noyes
sufficient specificity for clinical benefit
Imatinib and Related Drugs Target Philadelphia Chromosome Positive CML
bull BCR-ABL fusion creates a dys-regulated protein kinase that drives the proliferation of CML clones
protein kinase active site
bind ATP
bind target protein substrate
imatinib
specific kinase inhibition
A priori targeting the protein substrate binding site might be expected to provide more specificity buthelliphelliphelliphelliphelliphelliphellip
The Imatinib Experience
bull 82 of patients achieve a complete cytogenetic response (65-85)bull majority achieve a major molecular response (40-60)bull 7 year overall survival rate 90 bull event free survival rate 81 (65-85)
Positives
results from the International Randomized Study of Interferon Versus STI571 (IRIS) trial 7-year follow up
Blood (2008) 112 186
various studies from 2003-2008
bull Imatinib however does not destroy the cancer stem cell clone
bull If you remove imatinib CML returns
bull Imatinib is the perfect drug because once you are on it you are on it forever
If you place a cancer cell under selective pressure for long enough without killing it what will happen
- Slide 1
- Slide 2
- Slide 3
- Slide 4
- Slide 5
- Slide 6
- Slide 7
- Slide 8
- Slide 9
- Slide 10
- Slide 11
- Slide 12
- Slide 13
- Slide 14
- Slide 15
- Slide 16
- Slide 17
- Slide 18
- Slide 19
- Slide 20
- Slide 21
- Slide 22
- Slide 23
- Slide 24
-
GA101 is More Effective than Rituximab in Depleting B Cells in Clonus Monkeys
G R
Of Trumps Rituximab in a Clinical Trial for Chronic lymphocytic Leukemia
One Last Look at Where We would be Without Therapeutic Antibodies
Imatinib and Beyond Protein Kinase Inhibitors as Cancer Therapeutics
Protein Kinases as Drug Targets
Problem
There are over 500 protein kinases encoded by the human genome Can a drug target just one
Cancer Drug Gleevec (Imatinib) Wins Lasker AwardPosted at 343 pm CT on September 14 2009
The shortened Philadelphia chromosome seen in certain leukemias (picture from naturecom)The big science news of the day was the announcement of the Lasker Awards informally thought of as the American version of the Nobel Prize for physiology and medicine This yearrsquos clinical medical research award went to a trio of researchers from Oregon Health amp Science University Sloan-Kettering Cancer Center and drug company Novartis but you could just as easily say it was awarded to a drug the cancer treatment Gleevec (imatinib)
Noyes
sufficient specificity for clinical benefit
Imatinib and Related Drugs Target Philadelphia Chromosome Positive CML
bull BCR-ABL fusion creates a dys-regulated protein kinase that drives the proliferation of CML clones
protein kinase active site
bind ATP
bind target protein substrate
imatinib
specific kinase inhibition
A priori targeting the protein substrate binding site might be expected to provide more specificity buthelliphelliphelliphelliphelliphelliphellip
The Imatinib Experience
bull 82 of patients achieve a complete cytogenetic response (65-85)bull majority achieve a major molecular response (40-60)bull 7 year overall survival rate 90 bull event free survival rate 81 (65-85)
Positives
results from the International Randomized Study of Interferon Versus STI571 (IRIS) trial 7-year follow up
Blood (2008) 112 186
various studies from 2003-2008
bull Imatinib however does not destroy the cancer stem cell clone
bull If you remove imatinib CML returns
bull Imatinib is the perfect drug because once you are on it you are on it forever
If you place a cancer cell under selective pressure for long enough without killing it what will happen
- Slide 1
- Slide 2
- Slide 3
- Slide 4
- Slide 5
- Slide 6
- Slide 7
- Slide 8
- Slide 9
- Slide 10
- Slide 11
- Slide 12
- Slide 13
- Slide 14
- Slide 15
- Slide 16
- Slide 17
- Slide 18
- Slide 19
- Slide 20
- Slide 21
- Slide 22
- Slide 23
- Slide 24
-
Of Trumps Rituximab in a Clinical Trial for Chronic lymphocytic Leukemia
One Last Look at Where We would be Without Therapeutic Antibodies
Imatinib and Beyond Protein Kinase Inhibitors as Cancer Therapeutics
Protein Kinases as Drug Targets
Problem
There are over 500 protein kinases encoded by the human genome Can a drug target just one
Cancer Drug Gleevec (Imatinib) Wins Lasker AwardPosted at 343 pm CT on September 14 2009
The shortened Philadelphia chromosome seen in certain leukemias (picture from naturecom)The big science news of the day was the announcement of the Lasker Awards informally thought of as the American version of the Nobel Prize for physiology and medicine This yearrsquos clinical medical research award went to a trio of researchers from Oregon Health amp Science University Sloan-Kettering Cancer Center and drug company Novartis but you could just as easily say it was awarded to a drug the cancer treatment Gleevec (imatinib)
Noyes
sufficient specificity for clinical benefit
Imatinib and Related Drugs Target Philadelphia Chromosome Positive CML
bull BCR-ABL fusion creates a dys-regulated protein kinase that drives the proliferation of CML clones
protein kinase active site
bind ATP
bind target protein substrate
imatinib
specific kinase inhibition
A priori targeting the protein substrate binding site might be expected to provide more specificity buthelliphelliphelliphelliphelliphelliphellip
The Imatinib Experience
bull 82 of patients achieve a complete cytogenetic response (65-85)bull majority achieve a major molecular response (40-60)bull 7 year overall survival rate 90 bull event free survival rate 81 (65-85)
Positives
results from the International Randomized Study of Interferon Versus STI571 (IRIS) trial 7-year follow up
Blood (2008) 112 186
various studies from 2003-2008
bull Imatinib however does not destroy the cancer stem cell clone
bull If you remove imatinib CML returns
bull Imatinib is the perfect drug because once you are on it you are on it forever
If you place a cancer cell under selective pressure for long enough without killing it what will happen
- Slide 1
- Slide 2
- Slide 3
- Slide 4
- Slide 5
- Slide 6
- Slide 7
- Slide 8
- Slide 9
- Slide 10
- Slide 11
- Slide 12
- Slide 13
- Slide 14
- Slide 15
- Slide 16
- Slide 17
- Slide 18
- Slide 19
- Slide 20
- Slide 21
- Slide 22
- Slide 23
- Slide 24
-
One Last Look at Where We would be Without Therapeutic Antibodies
Imatinib and Beyond Protein Kinase Inhibitors as Cancer Therapeutics
Protein Kinases as Drug Targets
Problem
There are over 500 protein kinases encoded by the human genome Can a drug target just one
Cancer Drug Gleevec (Imatinib) Wins Lasker AwardPosted at 343 pm CT on September 14 2009
The shortened Philadelphia chromosome seen in certain leukemias (picture from naturecom)The big science news of the day was the announcement of the Lasker Awards informally thought of as the American version of the Nobel Prize for physiology and medicine This yearrsquos clinical medical research award went to a trio of researchers from Oregon Health amp Science University Sloan-Kettering Cancer Center and drug company Novartis but you could just as easily say it was awarded to a drug the cancer treatment Gleevec (imatinib)
Noyes
sufficient specificity for clinical benefit
Imatinib and Related Drugs Target Philadelphia Chromosome Positive CML
bull BCR-ABL fusion creates a dys-regulated protein kinase that drives the proliferation of CML clones
protein kinase active site
bind ATP
bind target protein substrate
imatinib
specific kinase inhibition
A priori targeting the protein substrate binding site might be expected to provide more specificity buthelliphelliphelliphelliphelliphelliphellip
The Imatinib Experience
bull 82 of patients achieve a complete cytogenetic response (65-85)bull majority achieve a major molecular response (40-60)bull 7 year overall survival rate 90 bull event free survival rate 81 (65-85)
Positives
results from the International Randomized Study of Interferon Versus STI571 (IRIS) trial 7-year follow up
Blood (2008) 112 186
various studies from 2003-2008
bull Imatinib however does not destroy the cancer stem cell clone
bull If you remove imatinib CML returns
bull Imatinib is the perfect drug because once you are on it you are on it forever
If you place a cancer cell under selective pressure for long enough without killing it what will happen
- Slide 1
- Slide 2
- Slide 3
- Slide 4
- Slide 5
- Slide 6
- Slide 7
- Slide 8
- Slide 9
- Slide 10
- Slide 11
- Slide 12
- Slide 13
- Slide 14
- Slide 15
- Slide 16
- Slide 17
- Slide 18
- Slide 19
- Slide 20
- Slide 21
- Slide 22
- Slide 23
- Slide 24
-
Imatinib and Beyond Protein Kinase Inhibitors as Cancer Therapeutics
Protein Kinases as Drug Targets
Problem
There are over 500 protein kinases encoded by the human genome Can a drug target just one
Cancer Drug Gleevec (Imatinib) Wins Lasker AwardPosted at 343 pm CT on September 14 2009
The shortened Philadelphia chromosome seen in certain leukemias (picture from naturecom)The big science news of the day was the announcement of the Lasker Awards informally thought of as the American version of the Nobel Prize for physiology and medicine This yearrsquos clinical medical research award went to a trio of researchers from Oregon Health amp Science University Sloan-Kettering Cancer Center and drug company Novartis but you could just as easily say it was awarded to a drug the cancer treatment Gleevec (imatinib)
Noyes
sufficient specificity for clinical benefit
Imatinib and Related Drugs Target Philadelphia Chromosome Positive CML
bull BCR-ABL fusion creates a dys-regulated protein kinase that drives the proliferation of CML clones
protein kinase active site
bind ATP
bind target protein substrate
imatinib
specific kinase inhibition
A priori targeting the protein substrate binding site might be expected to provide more specificity buthelliphelliphelliphelliphelliphelliphellip
The Imatinib Experience
bull 82 of patients achieve a complete cytogenetic response (65-85)bull majority achieve a major molecular response (40-60)bull 7 year overall survival rate 90 bull event free survival rate 81 (65-85)
Positives
results from the International Randomized Study of Interferon Versus STI571 (IRIS) trial 7-year follow up
Blood (2008) 112 186
various studies from 2003-2008
bull Imatinib however does not destroy the cancer stem cell clone
bull If you remove imatinib CML returns
bull Imatinib is the perfect drug because once you are on it you are on it forever
If you place a cancer cell under selective pressure for long enough without killing it what will happen
- Slide 1
- Slide 2
- Slide 3
- Slide 4
- Slide 5
- Slide 6
- Slide 7
- Slide 8
- Slide 9
- Slide 10
- Slide 11
- Slide 12
- Slide 13
- Slide 14
- Slide 15
- Slide 16
- Slide 17
- Slide 18
- Slide 19
- Slide 20
- Slide 21
- Slide 22
- Slide 23
- Slide 24
-
Protein Kinases as Drug Targets
Problem
There are over 500 protein kinases encoded by the human genome Can a drug target just one
Cancer Drug Gleevec (Imatinib) Wins Lasker AwardPosted at 343 pm CT on September 14 2009
The shortened Philadelphia chromosome seen in certain leukemias (picture from naturecom)The big science news of the day was the announcement of the Lasker Awards informally thought of as the American version of the Nobel Prize for physiology and medicine This yearrsquos clinical medical research award went to a trio of researchers from Oregon Health amp Science University Sloan-Kettering Cancer Center and drug company Novartis but you could just as easily say it was awarded to a drug the cancer treatment Gleevec (imatinib)
Noyes
sufficient specificity for clinical benefit
Imatinib and Related Drugs Target Philadelphia Chromosome Positive CML
bull BCR-ABL fusion creates a dys-regulated protein kinase that drives the proliferation of CML clones
protein kinase active site
bind ATP
bind target protein substrate
imatinib
specific kinase inhibition
A priori targeting the protein substrate binding site might be expected to provide more specificity buthelliphelliphelliphelliphelliphelliphellip
The Imatinib Experience
bull 82 of patients achieve a complete cytogenetic response (65-85)bull majority achieve a major molecular response (40-60)bull 7 year overall survival rate 90 bull event free survival rate 81 (65-85)
Positives
results from the International Randomized Study of Interferon Versus STI571 (IRIS) trial 7-year follow up
Blood (2008) 112 186
various studies from 2003-2008
bull Imatinib however does not destroy the cancer stem cell clone
bull If you remove imatinib CML returns
bull Imatinib is the perfect drug because once you are on it you are on it forever
If you place a cancer cell under selective pressure for long enough without killing it what will happen
- Slide 1
- Slide 2
- Slide 3
- Slide 4
- Slide 5
- Slide 6
- Slide 7
- Slide 8
- Slide 9
- Slide 10
- Slide 11
- Slide 12
- Slide 13
- Slide 14
- Slide 15
- Slide 16
- Slide 17
- Slide 18
- Slide 19
- Slide 20
- Slide 21
- Slide 22
- Slide 23
- Slide 24
-
Imatinib and Related Drugs Target Philadelphia Chromosome Positive CML
bull BCR-ABL fusion creates a dys-regulated protein kinase that drives the proliferation of CML clones
protein kinase active site
bind ATP
bind target protein substrate
imatinib
specific kinase inhibition
A priori targeting the protein substrate binding site might be expected to provide more specificity buthelliphelliphelliphelliphelliphelliphellip
The Imatinib Experience
bull 82 of patients achieve a complete cytogenetic response (65-85)bull majority achieve a major molecular response (40-60)bull 7 year overall survival rate 90 bull event free survival rate 81 (65-85)
Positives
results from the International Randomized Study of Interferon Versus STI571 (IRIS) trial 7-year follow up
Blood (2008) 112 186
various studies from 2003-2008
bull Imatinib however does not destroy the cancer stem cell clone
bull If you remove imatinib CML returns
bull Imatinib is the perfect drug because once you are on it you are on it forever
If you place a cancer cell under selective pressure for long enough without killing it what will happen
- Slide 1
- Slide 2
- Slide 3
- Slide 4
- Slide 5
- Slide 6
- Slide 7
- Slide 8
- Slide 9
- Slide 10
- Slide 11
- Slide 12
- Slide 13
- Slide 14
- Slide 15
- Slide 16
- Slide 17
- Slide 18
- Slide 19
- Slide 20
- Slide 21
- Slide 22
- Slide 23
- Slide 24
-
The Imatinib Experience
bull 82 of patients achieve a complete cytogenetic response (65-85)bull majority achieve a major molecular response (40-60)bull 7 year overall survival rate 90 bull event free survival rate 81 (65-85)
Positives
results from the International Randomized Study of Interferon Versus STI571 (IRIS) trial 7-year follow up
Blood (2008) 112 186
various studies from 2003-2008
bull Imatinib however does not destroy the cancer stem cell clone
bull If you remove imatinib CML returns
bull Imatinib is the perfect drug because once you are on it you are on it forever
If you place a cancer cell under selective pressure for long enough without killing it what will happen
- Slide 1
- Slide 2
- Slide 3
- Slide 4
- Slide 5
- Slide 6
- Slide 7
- Slide 8
- Slide 9
- Slide 10
- Slide 11
- Slide 12
- Slide 13
- Slide 14
- Slide 15
- Slide 16
- Slide 17
- Slide 18
- Slide 19
- Slide 20
- Slide 21
- Slide 22
- Slide 23
- Slide 24
-