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Is Ofatumumab Better than Rituximab? Ofatumumab is a fully human antibody produced in transgenic mice immunized with CD20 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) The trial did not meet its primary endpoint as there was no statistically significant difference in progression free survival (PFS) between the treatment arms. According to the firms, there were no differences in adverse events (AEs) leading to treatment discontinuation, Grade >3 AEs, severe adverse events (SAEs), or fatal SAEs between the treatment arms. "Based on today's results, we are unlikely to move forward with a regulatory filing," Winkel said.
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Page 1: 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
Page 2: BIOTECHNOLOGICAL APPLICATIONS 1Therapeutic antibodies 6_humanization

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
Page 3: BIOTECHNOLOGICAL APPLICATIONS 1Therapeutic antibodies 6_humanization

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
Page 4: BIOTECHNOLOGICAL APPLICATIONS 1Therapeutic antibodies 6_humanization

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
Page 5: BIOTECHNOLOGICAL APPLICATIONS 1Therapeutic antibodies 6_humanization

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
Page 6: BIOTECHNOLOGICAL APPLICATIONS 1Therapeutic antibodies 6_humanization

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
Page 7: BIOTECHNOLOGICAL APPLICATIONS 1Therapeutic antibodies 6_humanization

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
Page 8: BIOTECHNOLOGICAL APPLICATIONS 1Therapeutic antibodies 6_humanization

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
Page 9: BIOTECHNOLOGICAL APPLICATIONS 1Therapeutic antibodies 6_humanization

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
Page 10: BIOTECHNOLOGICAL APPLICATIONS 1Therapeutic antibodies 6_humanization

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
Page 11: BIOTECHNOLOGICAL APPLICATIONS 1Therapeutic antibodies 6_humanization

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
Page 12: BIOTECHNOLOGICAL APPLICATIONS 1Therapeutic antibodies 6_humanization

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
Page 13: BIOTECHNOLOGICAL APPLICATIONS 1Therapeutic antibodies 6_humanization

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
Page 14: BIOTECHNOLOGICAL APPLICATIONS 1Therapeutic antibodies 6_humanization

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
Page 15: BIOTECHNOLOGICAL APPLICATIONS 1Therapeutic antibodies 6_humanization

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
Page 16: BIOTECHNOLOGICAL APPLICATIONS 1Therapeutic antibodies 6_humanization

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
Page 17: BIOTECHNOLOGICAL APPLICATIONS 1Therapeutic antibodies 6_humanization

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
Page 18: BIOTECHNOLOGICAL APPLICATIONS 1Therapeutic antibodies 6_humanization

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
Page 19: BIOTECHNOLOGICAL APPLICATIONS 1Therapeutic antibodies 6_humanization

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
Page 20: BIOTECHNOLOGICAL APPLICATIONS 1Therapeutic antibodies 6_humanization

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
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  • Slide 19
  • Slide 20
  • Slide 21
  • Slide 22
  • Slide 23
  • Slide 24
Page 21: BIOTECHNOLOGICAL APPLICATIONS 1Therapeutic antibodies 6_humanization

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
Page 22: BIOTECHNOLOGICAL APPLICATIONS 1Therapeutic antibodies 6_humanization

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

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Page 23: BIOTECHNOLOGICAL APPLICATIONS 1Therapeutic antibodies 6_humanization

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

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Page 24: BIOTECHNOLOGICAL APPLICATIONS 1Therapeutic antibodies 6_humanization

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

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  • Slide 2
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