Surrobodies™ – A Novel Approach to Bispecifics… Lane IBC DDT deck 080612.pdf · 6/12/2008 ·...
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Transcript of Surrobodies™ – A Novel Approach to Bispecifics… Lane IBC DDT deck 080612.pdf · 6/12/2008 ·...
Surrobodies™ – A Novel Approach to Bispecifics…
Ramesh Bhatt, Ph.D.Vice President of ResearchSea Lane Biotechnologies
1Sea Lane – DDD presentation August 8, 2012 Sea Lane Biotechnologies – Proprietary
Presentation Agenda
Introduction to Surrobody structureSurrobody advantages directly address bispecific challengesThree brief bispecific examples
Comparability to antibodiesInhibiting soluble targets in vitro and in vivoTargeting receptors in vitro
SummarySurrobody Drug Conjugate Teaser
2Sea Lane – DDD presentation August 8, 2012 Sea Lane Biotechnologies – Proprietary
SurrobodiesHighly adaptable platform inspired by
endogenous human pre-B Cell Receptor
SurrogateLight Chain
VpreB
λ5
VpreB
λ5
Fused SLCNative SLC
3Sea Lane – DDD presentation August 8, 2012 Sea Lane Biotechnologies – Proprietary
The Sea Lane Advantage
• The invariant surrogate light chain simplifiesand accelerates bispecific development and enables drug conjugation for all existing and future Surrobodies
• Proprietary design of fully human, synthetic libraries mirror natural human diversity enabling rapid, high quality lead generation and simple, comprehensive optimization
• Use of naturally occurring, human germline frameworks provides consistently robust expression characteristics and should reduce immunogenic potential
Surrobody™
SurrogateLight Chain
(blue/yellow)
HeavyChain
(green)
4Sea Lane – DDD presentation August 8, 2012 Sea Lane Biotechnologies – Proprietary
Surrogate Light Chain Only known universal heavy chain partner Expected low immunogenicity Invariant protein that simplifies all steps from discovery through manufacture
Intact FcImparts effector function and favorable half lifeKnown scalable production capable through Protein A
Bispecific/Drug Conjugate ReadyRapid conversion to enhanced format
Intellectual PropertyGranted patents protect basic structure and synthetic human library design
General Surrobody Strengths
Complete Heavy Chain
SLC
5Sea Lane – DDD presentation August 8, 2012 Sea Lane Biotechnologies – Proprietary
Synthetic Human Surrobody LibraryDrives Discovery
~28 billion productive membersPatented human diversity productively matches frameworks and CDRs
Based on functional diversityPowers companion optimization technology
Novel library construction processes maximize library fidelity12 week panning through lead ID cycle timeSurrobodies found against 14 different therapeutic targets
6Sea Lane – DDD presentation August 8, 2012 Sea Lane Biotechnologies – Proprietary
Surrobodies Provide Ideal Path for Bispecific and Drug Conjugate Applications
BispecificsSimplified structure dramatically reduces required resourcesAntibody-like PK and other beneficial CMC qualitiesRobust in vivo efficacyMultiple bispecific formats provide unmatched flexibility
Toxin Drug ConjugationSimplified process utilizing the universal surrogate light chain is applicable to all SurrobodiesBioanalytic assessment supports robust and stable conjugation Resulting toxin conjugated Surrobodies are highly potent
7Sea Lane – DDD presentation August 8, 2012 Sea Lane Biotechnologies – Proprietary
Fc Engineering Improves Bispecific Assembly but Provides an Incomplete Solution
25% Productive
75% Unwanted
A A B B
BA
BABA BA
Fc engineered heteromericHeavy chain assembly
8
Light chain mismatching generates predominant and undesirable products
Sea Lane – DDD presentation August 8, 2012 Sea Lane Biotechnologies – Proprietary
Sea Lane Advantage:The Surrogate Light Chain
9
Invariant surrogate light chain partnering eliminates unproductive combinations caused by mismatching of light chain/heavy chain partners
A A
BA
B B
Sea Lane – DDD presentation August 8, 2012 Sea Lane Biotechnologies – Proprietary
Bispecific Example #1Comparability to Antibodies
10Sea Lane – DDD presentation August 8, 2012 Sea Lane Biotechnologies – Proprietary
Surrobodies DisplayAntibody-Like Characteristics
BiochemicalLong term RT stability >1yrHigh level protein expression
Many >100mg/L in HEK293
Stable following physical stressfreeze/thaw cyclesLow pH 3 exposureShort term High temp exposure
• at 50°C for 48 hours
High concentration (100mg/ml)
All Surrobody formats maintain thermal stability profiles that are similar to commercial antibodies
Tm >65°C
In vitroHigh affinity binding (subnanomolar)Antagonist and Agonist activities are possibleBroad epitopic range
In VivoNHP half-life ~1-2 wksExcellent efficacy in rodent models of disease
11Sea Lane – DDD presentation August 8, 2012 Sea Lane Biotechnologies – Proprietary
Monovalent Bispecifc Surrobodies Simultaneously Bind Two Distinct Targets
0
1
2
3
4
0.01 0.1 1 10 100
OD
450
nm
[nM]
HGF + PlGF :Fc(bi)
P
H
HGF
BiotinPlGF
SAHRP
12
HGF + PlGFBispecific
Sea Lane – DDD presentation August 8, 2012 Sea Lane Biotechnologies – Proprietary
Mammalian Expressed Surrobodies Have Excellent Characteristics
Format
Unique proteins(# preps)
Avg yield
(mg/L)
Intact SgG (%)
High MW“Aggregate”
(%)
Low MW“Protein”
(%)
Thermal Stability
(oC)
RT Stability (months)
Standard Surrobody
194(249)
77 98.2% 1.6% 0.2% >65 24
Bispecific Monovalent
Binding
72(82)
81 88.2% 3.4% 8.4% >65 12
All 16 possible bispecific Vh framework combinations produce comparable yields in HEK293 transient systems
13Sea Lane – DDD presentation August 8, 2012 Sea Lane Biotechnologies – Proprietary
Size Exclusion and Mass Spec Show High Quality Monovalent Bispecific Complexes
mass147000 148000 149000 150000
%
0
100 94.8147981.1
148072.1
SL-183
Bi-specificTarget 1:Target 2Mass Spec analysis:
Virtually all heterodimeric bispecific
SEC analysis: Cotransfection results in single peak at the appropriate size marker
MWStandards
Protein A purifiedBispecific Surrobody
14Sea Lane – DDD presentation August 8, 2012 Sea Lane Biotechnologies – Proprietary
7400 147600 147800 148000 148200 148400 148600 1488
7_sample05 157 (5.321) M1 [Ev-120888,It10] (Gs,0.300,2207:3792,0.30,L30,R30); Cm (154:167)148102.4
148201.7
147400 147600 147800 148000 148200 148400
196 (6.641) M1 [Ev-147837,It10] (Gs,0.300,2015:3843,0.30,L30,R30); Cm (193:205)147800.9
147899.0
147400 147600 147800 148000 148200 148400
184 (6.234) M1 [Ev-142519,It10] (Gs,0.300,2042:3846,0.30,L30,R30); Cm (179:191)147844.1
147942.8
148041.8
147400 147600 147800 148000 148200 148400 148600
_sample08 166 (5.625) M1 [Ev-141161,It10] (Gs,0.300,2158:3753,0.30,L30,R30); Cm (159:182)147884.6
147984.8
147400 147600 147800 148000 148200 148400 1486
87_sample09 206 (6.979) M1 [Ev-140978,It10] (Gs,0.300,2014:3660,0.30,L30,R30); Cm (201:218)147888.8
147987.5
147700 147800 147900 148000 148100 148200 148300
] ( ) ( )148004.5938
148104.2031
7400 147600 147800 148000 148200 148400 148600
118) M1 [Ev-120766,It10] (Gs,0.300,2156:3427,0.30,L30,R30); Cm (145:171)148013.5938
147588.5000
148112.0000
7400 147600 147800 148000 148200 148400 148600
03 190 (6.437) M1 [Ev-158758,It10] (Gs,0.300,2023:3934,0.30,L30,R30); Cm (187:206)148017.5000
148116.7969
147400 147600 147800 148000 148200 148400 148600 148800
175 (5.930) M1 [Ev-146679,It10] (Gs,0.300,2137:4000,0.30,L30,R30); Cm (172:189)148061.0000
148158.7969
H/P H/V H/B5 H/B6
H/D V/B5 V/B6 V/D B5/D
Deglycosylated proteins
(Focusing on region of intact tetramers:
~148,000 Da)
LC-MS Analysis of a Collection of Monovalent Bispecific Surrobodies
15Sea Lane – DDD presentation August 8, 2012 Sea Lane Biotechnologies – Proprietary
Monovalent Bispecific Surrobodies are Stable Following Storage at Room Temp for 12 Months
ELISA binding isotherms show no loss of activity between -80°C and room temperature.
-1 0 1 2 30
1
2
3
4-80oC4oCR.T
log [conc] nM
OD
450n
m
EC50 (nM)
-80°C 1.92
4°C 2.12
Room Temp 1.87
-80°C4°CR.T
16Sea Lane – DDD presentation August 8, 2012 Sea Lane Biotechnologies – Proprietary
The PK Properties of Bispecific Surrobodies are Comparable to Parentals in Cynomolgus
AgentHalf-life(days)
Clearance(mL/hr/kg)
Vss(mL/kg)
Bispecific (HGF/PlGF) 6.04 0.42 77.2
Monospecific PlGF 6.35 0.35 78.5
Monospecific HGF 9.28 0.40 110
HGF IgG 10.88 0.19 75.6
0 10 20 300.1
1
10
100
1000 Bispecific HGF/PlGF
Monospecific PlGFMonospecific HGF
time (days)
ug/m
l
Naïve groups (n=3) of Cynomolgus monkeys were administered single IV dose of Surrobodies (10 mg/kg)Serum was tested over a 28 day periodThe overall PK properties are “Antibody-like”
17Sea Lane – DDD presentation August 8, 2012 Sea Lane Biotechnologies – Proprietary
Surrobody Thermostability is Comparable to Marketed Antibodies
SL-0
02
SL-0
12
SL-1
76
SL-1
86
SL-1
88
Erbi
tux
Her
cept
in50
55
60
65
70
Tm° C
Target Format
SL-002 PlGF
MonospecificSL-012 HGF
SL-176 ErbB3
SL-186PlGFHGF
Bispecific v1
SL-188 Bispecific v2
Tm was calculated in a fluorescent-based assay of agents in PBS
18Sea Lane – DDD presentation August 8, 2012 Sea Lane Biotechnologies – Proprietary
Bispecific Example #2Targeting Two Soluble Factors
in vitro and in vivo
19Sea Lane – DDD presentation August 8, 2012 Sea Lane Biotechnologies – Proprietary
GF x Ligand Bispecific Properties are Similar to Parental Surrobodies
All values [nM]Ligand-1 Ligand-2 GF
Molecule(specificity)
CellIC50
ELISA Binding
ELISA Inhibition
ELISA Binding
CellIC50
SL-429(Ligand) 0.281 0.020 0.156
SL-516(GF) 0.086 0.139
SL-634(Ligand x GF)MonovalentBispecific
Not Tested 0.048 0.955 0.055 2.602
20Sea Lane – DDD presentation August 8, 2012 Sea Lane Biotechnologies – Proprietary
Anti-GF/Ligand Bispecific Captures the Synergistic Benefits of Dual Inhibition In Vivo
Mice were treated 2x weekly by iv injection. Groups received 5mg/kg each, except bispecific which was 10mg/kg
21Sea Lane – DDD presentation August 8, 2012
Colo205 Day 54
0
500
1000
1500
2000
100%TGI
73%
α-ligand
58%
α-GF
94%
α-GF + α-Ligandcocktail
96%
α-Ligand/α-GFBispecific
Median tumorgrowth inhibition
0%
PBS
Tum
or v
olum
e (m
m3 )
+
Sea Lane Biotechnologies – Proprietary
Anti-GF/Ligand Bispecific Captures the Synergistic Benefits of Dual Inhibition In Vivo
Colo205 Xenograft
1 8 15 22 29 36 43 50 570
500
1000
1500
2000
PBS
Anti-LigandAnti-GF (n = 9)
Anti-GF/Ligand BispecificAnti-GF + Anti-Ligand
Day of study
Mea
n tu
mor
vol
ume
(mm
3 ) ± S
EM
Mice were treated 2x weekly by iv injection. Groups received 5mg/kg each, except bispecific which was 10mg/kg
22Sea Lane – DDD presentation August 8, 2012 Sea Lane Biotechnologies – Proprietary
Bispecific Example #3In vitro Targeting
ErbB3 x Growth Factor Receptor
23Sea Lane – DDD presentation August 8, 2012 Sea Lane Biotechnologies – Proprietary
ErbB3 Summary
ErbB3 represents a pivotal node in regulation of cell proliferation
Implicated in the generation of resistance to a number of key oncology agents
Sea Lane’s anti-ErbB3 is a potent inhibitor with a novel mechanism of action in ErbB2 overexpressing cells (Molecular Cancer Therapeutics; July, 2012)
Sea Lane’s anti-ErbB3 Surrobodies synergistically enhance the activity of ErbB2 and EGFR antibodies
Bispecific ErbB3 x GFR surrobodies capably capture a similarly synergistic cocktail
24Sea Lane – DDD presentation August 8, 2012 Sea Lane Biotechnologies – Proprietary
Anti-ErbB3 Surrobodies Enhance the Activity of Trastuzumab
-4 -2 0 2
0
20
40
60
80
log [SgG/IgG] nM
Perc
ent I
nhib
ition
-4 -2 0 2
0
20
40
60
80
log [SgG/IgG] nM
Perc
ent I
nhib
ition SL-175
Trastuzumab
SL-175*+Trastuzumab
Pertuzumab+Trastuzumab
Pertuzumab
Unstimulated NRG-stimulated
* Nearly identical results were obtained using SL-176Foreman, et. al., Molecular Cancer Therapeutics 2012
SKBR3 Cells
25Sea Lane – DDD presentation August 8, 2012
Anti-ErbB3 Surrobody enhancement of anti-ErbB2 provides the rational basis for an ErbB3 x ErbB2 bispecific
Sea Lane Biotechnologies – Proprietary
Anti-ErbB3 Surrobodies Enhance the Activity of Cetuximab
-2 -1 0 1 2 3
0
50
100
SL-175+CetuximabSL-176+CetuximabCetuximab
SL-175SL-176
log [SgG/IgG] nM
Perc
ent I
nhib
ition
Anti-ErbB3 Surrobody enhancement of anti-EGFR provides the rational basis for an ErbB3 x EGFR bispecific
A431 cells
Sea Lane Biotechnologies – Proprietary 26Sea Lane – DDD presentation August 8, 2012
Bispecific ErbB3 x GFR Surrobody Captures Complimentary Efficacy of Parental Cocktail
Anti-GFR ( ) Surrobody combined with anti-ErbB3 ( ) Surrobody
SL-433 (Monovalent bispecific)
0.01 0.1 1 10 100 1000
0
20
40
60
80
SL-176 (Anti-ErbB3)
SL-396 (Anti-GFR)
SL-396 + SL-176
[SgG] nM
Per
cent
Inhi
bitio
n
SL-433
27Sea Lane – DDD presentation August 8, 2012 Sea Lane Biotechnologies – Proprietary
Summary
The Surrogate Light Chain is a natural universal Heavy Chain partner
Sea Lane’s fully human synthetic library drives Surrobody discovery
High level, high quality transient expression accelerates Surrobodies through in vitro and in vivo research
Bispecific Surrobodies are highly efficacious with characteristics that are comparable to antibodies
The Surrobody structure is amenable to additional formats that enable optimal therapeutic approaches including:
Multivalent bispecifics
Drug conjugation
28Sea Lane – DDD presentation August 8, 2012 Sea Lane Biotechnologies – Proprietary
Surrobody Drug Conjugates
Boosting the anti-tumor abilities of Surrobodies
29Sea Lane – DDD presentation August 8, 2012 Sea Lane Biotechnologies – Proprietary
Surrobody Drug Conjugates
Invariant surrogate light chain partner is the ideal site for drug conjugation
ALL Surrobodies use the exact same light chain
One solution will work across the entire platform from monospecific to bispecific agents
Light chain has been shown to be more desirable for conjugation than heavy chain
30Sea Lane – DDD presentation August 8, 2012 Sea Lane Biotechnologies – Proprietary
Surrobody Drug ConjugationResearch Milestones
Engineered Surrobody for drug conjugationModified surrogate light chain allows for 2 toxins per SurrobodyPossible to further modify to add more toxinToxin addition efficiency appears excellent
Demonstrated cellular potencies in vitroPOC target: GFR driven cell proliferation
Bispecific drug conjugation comparable to monospecific Surrobodies
31Sea Lane – DDD presentation August 8, 2012 Sea Lane Biotechnologies – Proprietary
Toxins Conjugate Robustly to Surrobodies
Modified Surrobody w/o toxinModified Surrobody plus toxin
0 1 2drugs per molecule
• Analysis of unoptimized Drug Conjugation shows predominance of Surrobodies containing two or one toxins per molecule
• Toxin conjugate: MonomethylAuristatin E (MMAE)
32Sea Lane – DDD presentation August 8, 2012 Sea Lane Biotechnologies – Proprietary
Possible to modify to add more toxinsPerforms as well as, or better than toxin conjugated antibody (not shown)
Surrobody Drug Conjugates More Potently Inhibit In Vitro Proliferation
33Sea Lane – DDD presentation August 8, 2012
0.0001 0.001 0.01 0.1 1 10 100 1000
0
20
40
60
80
100
Parent SurrobodyEngineered SLC controlEngineered SLC #1 + ConjugateEngineered SLC #2 + Conjugate
[IgG/SgG] in nM
per
cen
t in
hib
itio
n
Sea Lane Biotechnologies – Proprietary
Potency[pM] Efficacy
Parent 0.615 20%
Engineered SLC control 0.382 19%
Engineered SLC #1 + Conjugate
0.058 96%
Engineered SLC #2 + Conjugate
0.156 99%
+1 +20Drug permolecule
“A” x “B”
+1 +20Drug permolecule
“A” x “C”
+1 +20Drug permolecule
“B” x “C”
Blue tracing is Unmodified SgG/ Red tracing is toxin (MMAE) conjugated
Bispecific Surrobody Toxin Conjugation is Comparable to Monospecific Conjugation
Heteromeric Fc Bispecific Surrobodies
+1 +20Drug permolecule
“A” x “N”
+1 +20Drug permolecule
“N” x “A”
+1 +20Drug permolecule
“B” x “N”
In unoptimized conjugation reactions almost all the surrobodies are conjugated with most carrying two toxin molecules
34Sea Lane – DDD presentation August 8, 2012 Sea Lane Biotechnologies – Proprietary
Acknowledgements
Technology DevelopmentLi XuSandra WangGordon Leung
Protein SciencesChuck HannumHieu Tran
DiscoveryAaron KurtzmanHelena YeeDavid Duffin
Program TeamsDanying CaiMedini GorePamela ForemanPhil Kobel
ManagementLawrence HorowitzMichael Horowitz
35Sea Lane – DDD presentation August 8, 2012 Sea Lane Biotechnologies – Proprietary
Contact Information
Sea Lane Biotechnologies2450 Bayshore Parkway, Suite 200Mountain View, CA 94043USAMichael Horowitz Ramesh BhattGC/COO VP of [email protected] [email protected](650) 336-1532 (650) 325-7402
36Sea Lane – DDD presentation August 8, 2012 Sea Lane Biotechnologies – Proprietary