OmniAb® · PDF file · 2017-10-04Evaluate Pharma February 2016 0 2 4 6 8 10 12...
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Transcript of OmniAb® · PDF file · 2017-10-04Evaluate Pharma February 2016 0 2 4 6 8 10 12...
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Antibodies, Leading Therapeutic Class
Evaluate Pharma February 2016
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Hum
ira (T
NF)
Revlim
id
Opd
ivo (PD‐1)
Harvo
ni
Prevna
r
Avastin
(VEG
F)
Hercerptin
(Her2)
Soliris (C
5)
Tecfidera
Orkam
bi
Entresto
Rituxan (CD20
)
Enbrel (T
NF)
Remicad
e (TNF)
Xtan
di
Janu
via
Keytruda
(PD‐1)
Eliquis
Eylea (VEG
F)
Triumeq
Top-20 product forecast, 2015-2020- Half antibodies (blue)- Half non-antibodies (orange)- Sales growth from $91B to $124B- 53% of sales from antibodies
Global sales ($B)
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$80B 2015E$150B 2020E
Murine (5) Chimeric (9) Humanized (33) Human phage (5) Human transgenic (19) Human (1)
ReviewRomosozumab (UCB)Inotuzumab (Pfizer)Ocrelizumab (Roche)
Sirukumab (Janssen)Dupilumab (Regeneron)Brodalumab (Amgen)Sarilumab (Regeneron)
Xilonix (Xbiotech)
2016Obiltoxaximab (Elusys)Infliximab EU (Bioepis)Infliximab US (Celltrion)
Elotuzumab (BMS)Mepolizumab (GSK)Atezolizumab (Roche)Reslizumab (Teva)Ixekizumab (Lilly)
Olaratumab (Lilly)Bezlotoxumab (Merck) Daratumumab (Janssen)Necitumumab (Lilly)
2015 Dinutuximab (United) Idarucizumab (BI)Mepolizumab (GSK) Necitumumab (Lilly)
Alirocumab (REGN)Evolocumab (Amgen)Secukinumab (Novartis)
2014 Blinatumomab (Amgen) Siltuximab (Janssen)Pembrolizumab (Merck)Vedolizumab (Takeda)Numax (AZ)
Adalimumab IN (Cadila) Nivolumab (BMS)
2013 Infliximab EU (Celltrion)
Trastuzumab Emt. (Roche)Obinutuzumab (Roche)Mogamulizumab (Kirin)Trastuzumab IN (Biocon)Itolizumab IN (Biocon)Adotrastuzumab (Roche)
Ramucirumab (Lilly)
2012 Pertuzumab (Roche) Raxibacumab (GSK)
2011 Brentuximab vedotin (SGEN) Ipilimumab (BMS)
00sCatumaxomab (Fresenius)I‐131 Tositumomab (GSK)Ibritumomab Tiu. (Biogen)
Cetuximab (Lilly)
Tocilizumab (Roche)Certolizumab Pegol (UCB)Eculizumab (Alexion)Ranibizumab (Roche)Bevacizumab (Roche)Natalizumab (Biogen)Nimotuzumab (Daiichi)Efalizumab (Roche)Omalizumab (Roche)Alemtuzumab (Sanofi)Gemtuzumab (Pfizer)
Belimumab (GSK)Adalimumab (AbbVie)
Denosumab (Amgen)Golimumab (Janssen)Ustekinumab (Janssen)Canakinumab (Novartis)Ofatumumab (GSK)Panitumumab (Amgen)
90s
Infliximab (Janssen)Basiliximab (Novartis)Rituximab (Roche)Abciximab (Janssen)
Palivizumab (AZ)Trastuzumab (Roche)Zenapax (Roche)
80s OKT3 (Janssen)
Reichert Oct 2016
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US antibody phase IIIs – Doubling in 5 years
26 25 25
29
33
39
53
0
10
20
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2010 2011 2012 2013 2014 2015 2016
Reichert Nov 2015
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US antibody clinical programs
90115
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150 115
17
0
50
100
150
200
250
300
Phase I Phase II Phase III
Non‐cancer
Cancer
Reichert Oct 2016
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Therapeutic antibody programs
1,021preclinical
342phase I
276phase II
137phase III
15 filing
49 distinct generic names
(including combos & company sharing)
BioPharm Insight August 2015
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OmniAb Advantage
OmniRat– Industry‐only rat– Freedom‐to‐operate– Industry‐first head‐to‐
head with wildtypeOmniMouse– Industry‐only dual species– Freedom‐to‐operate– No Regeneron law suitOmniFlic– Only rat for bispecifics– Freedom‐to‐operate– TeneoSeek avoids
hybridoma limitations
OmniAb– All non‐confidential info– www.omniab.comOptions– Front‐ or back‐loaded– Royalty ranges– Rat‐ or mouse‐onlyPartner control– Indications– Territories– Combinations– SublicensingAgreement– Easy 20 pages– Partner proven
Animal hosting– Global master colonies– All species and genetic makeupsOmniDeep– Gateway to diverse CROs– www.omnideep.com
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OmniAb “multidimensional advantage”
Scarce therapeutic targets 1 2 3 . . . n
Single monoclonal technology • Single‐platform epitope coverage, affinity, specificity, etc.
• Suboptimal antibody discovery for each scarce clinical target
OmniAb
• OmniRat• OmniMouse• OmniFlic
• Multiple species and genetic backgroundso Broader epitope coverageo Higher affinityo More specificity choices
• Bispecific antibodieso Simultaneously address two therapeutic targetso Engage effector functionso Strong intellectual property
• Greater antibody discovery for each scarce clinical target
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OmniDeep™
Industry‐only rat for
monospecific hmAb discovery
One of few mouse hmAb platforms with
FTO
OmniDeep™Global network of antibody
discovery CROs, hybridoma‐free
Industry‐only rat for
bispecific hmAb discovery
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OmniAb access options
• Platform access• Clinical milestones• Royalties
Unlimited license
• Target fee• Clinical milestones• Royalties
Individual target license
• Buy‐out fee• Single‐ or multi‐year• NO milestones or royalties
Unlimited buy‐out
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Partners29 user groups– 24 with unlimited access for milestones and/or royalties– 3 fully paid up in single transaction– 2 academics
Strategic partners– Three animal breeders and knock‐out providers– Four CROs ‐ US, Europe and Japan
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>100 successful antibody campaigns– >10,000 fully human high affinity binders
Good manufacturability, high affinity, expected PKFirst Phase I trial in 20153 INDs in 20165‐10 INDs in 2017
OmniAb Antibody Development
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Platforms for hmAb discovery
VH
CH3
CH2
hinge
VL
VH C
CH1
CH2
CH3
hinge
Ligand unlimited platform license TENEO sequence license
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Platform development
Inactivation of endogenous rat Ig genes– Heavy chain J‐locus– Light chain Cκ– Light chain Cλ
Recombinant immunoglobulin loci– Kappa light chain– Lambda light chain– Heavy chain
Improved genetic engineering based on proven technology
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Novel and proprietary knockout technology
Sangamo Zn‐finger technology– Exclusive license for Rat Ig knockout
Zn‐finger nuclease– One cut per genome– DSB repair– Mutation
Inactivation of rat antibody expression
Target sequence (exon of coding gene)
Non Homologous End‐Joining(NHEJ)
deletion insertion
Gene Disruption
DSB
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Micro‐injection, ZNF‐targeted mutagenesis
X
One‐cell embryoExtract DNA to look for ZFN activity
ZFN1/ZFN2
Transfer to pseudopregnant
females
Newborns
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Science‐first rat immunology gene knock‐out
Science2009, July 24, 325: 433‐
European Journal of Immunology2010, 40: 2932–2941
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OmniRat/OmniFlic/UniRat
New KO Technology–Heavy chain J‐locus deletion–Kappa light chain mutation–Lambda constant region deletion
Rat antibody expression 100% inactivated
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Transgenic immunoglobulin lociRepresenting the human V(D)J repertoire
m‐RNA Human Antibody
Human LC locus
VL…………… VL1 J Ck +
VJ C
VH………VH1 D J E Cm Cd C2b E A
Easy
Conversion
Heavy chain locus
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Transgenic immunoglobulin lociRepresenting the human V(D)J repertoire
m‐RNA Human Antibody
Human LC locus
VL…………… VL1 J Ck +
VJ C
VH………VH1 D J E Cm Cd C2b E A
Easy
Conversion
Heavy chain locus
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Rat and mouse transgenesis
X
One‐cell embryo
Microinject DNA construct
Implant in hormone‐treated female
Transgenic offspring
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OmniRat and OmniMouseFunctional recombinant immunoglobulin loci– Productive rearrangement of all functional human VH, DH, JH and VL, JL– Normal human frequencies of V‐, D‐, J‐gene usage– Normal human CDR3 length
Normal B‐cell developmentHigh expression of human antibodiesNormal hypermutation and affinity maturation
• Sprague Dawley• Brown Norway• Lewis
• B6/SJL
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ImmunizationStandard (every 3‐4 weeks +/‐ adjuvant)Rapid (2x/week for 4 weeks)Rapid (1x at base of the tail)DNA prime/protein boostCell prime/DNA boostAddition of T‐cell epitope
• Sprague Dawley• Brown Norway• Lewis
• B6/SJL
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Two species = more antibodies = better epitope coverage
Different antibody titers– Different immune response genes
• SD vs BN vs LEW vs Mouse Bl6/SJL
– Human antigen ≠ rat antigen ≠ mouse antigen– Different V‐genes (human vs rat vs mouse)
• Blocking Antibodies: OmniMouse vs OmniRat vs Mouse vs Rat
– Isotype Switching• Increased IgM+/decreased IgG+ B‐cells in OmniRat• Mouse vs Rat Fc
RatMouse
Epitope coverage
Gene Human/Mouse Human/Rat Mouse/RatCD30* 54.0% 50.1% 83.4%CD22* 58.7% 56.9% 77.7%CD14 63.7% 61.3% 80.9%CD80 39.2% 43.4% 63.4%CD52 36.1% 41.0% 64.9%
IL‐1 beta 64.7% 63.8% 86.9%
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Hybridoma generationOMT rats make antibodies as well as normal rats
Animal Antigen Cells* fusions titer hybrids IgGs** Kd***
SD PG LN 1 38400 3520 38 0.3‐1.0 nM
OmniRat PG LN 1 12800 1600 148 0.7‐2.4 nM
OmniRat hGHR LN 3 4800 704‐1024 18, 3, 2 ND
SD TAU/KLH LN 1 20000 1728 99# 0.6‐2.4 nM
OmniRat TAU/KLH LN 1 4800 1880 118# 0.5‐3.2 nM
SD HEL LN 1 12800 1564 26 0.02‐0.1 nM
OmniRat HEL LN 3 25600 288‐640 0, 2, 7 0.6‐1.5 nM
SD OVA LN 1 9600 1488 10 1.1‐4.8 nM
OmniRat OVA LN 4 8000 512‐2240 0, 30, 0, 1 0.7‐1.5 nM
5 different antigens
Single immunization on day 0Lymph node fusion on day 21
16 fusions
Similar titers
Similar # of hybridomas
502 mAbs confirmed by Biacore
5 highest affinity Abs
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High number of unique binders
Target Homology human/rat
# screened clones
# binders # unique binders
1 99% 7896 511 1752 98% 6768 337 1303 67% 2880 297 49
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Good diversity against conserved targets
20 IGHV, 11 IGKV, and 12 IGLV different germ lines isolated from 3 screening campaigns
Some V genes are specific to one target selection
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OmniRat vs phage display antibody binding
Target 1
* Estimated affinities
Target 2
Target 3
KD*= 1.9 nM
OMT‐A1
KD= 2.3 nM
KD= 15 nM KD= 0.01 nM
OMT‐C2
KD*= 0.001 nM KD*= 0.02 nM
OMT‐B1
OMT‐C3
KD= 7 nM
OMT‐C4
KD*= 4 nM
OMT‐C1
KD= 0.07 nM
KD*= 0.13 nM
OMT‐B2
KD= 0.33 nM
OMT‐B3
OMT‐A2
KD= 219 nM
Phage display‐derived IgG
OmniRat‐derived IgG
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DNA immunization
MONOCLONALPOLYCLONAL
Polyclonal sera (8 weeks) Monoclonal antibodies(4 months)
No need for isolated protein or peptide during the process or for screening
Start from electronic cDNA sequences
Cloning of human CEACAM5 cDNA into our specially designed
vectors
cDNA immunisation (GENOVAC Antibody Technology®) of
OmniRat and Wistar rats. Immune
response against the native antigen
Fusion and hybridoma generation
Screening and sub cloning
Serum and supernatant test on native protein. Cross‐reactivity tests.
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Receptor protein complex
Immune serum (1:1000 dilution) of a representative animal is tested on
mammalian cells transfected with the cDNA encoding for the target antigen
(green curves) or with an irrelevant construct (red curves)
Three fusions with 7 immunized animals– 505 positive hits out of 960 tested samples (53%)– 1122 positive hits out of 1632 tested samples (69%)– 792 positive hits out of 864 tested samples (69%)
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GPCR
Immune serum (1:1000 dilution) of a representative animal is tested on
mammalian cells transfected with the cDNA encoding for the target antigen
(human = green curves, mouse = blue), on a stable cell line, or with an irrelevant
construct (red curves)
Parallel immunization with KO mice unsuccessful
Three fusions with 10 immunized animals– 11 positive hits out of 1824 tested samples (0.6%)– 34 positive hits out of 1920 tested samples (1.8%)– 2 positive hits out of 1920 tested samples (0.1%)
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Transgenic animals for hmAb discovery
Fixed L‐chain IgG antibodies
OmniFlic with rearranged human L‐chain expressed with any (human) IgH locus
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Immune response – cellular perspectiveMultiple injections, 5‐6 week immunization time course Harvest cells from lymph nodes
Millions of naïveB cells in circulation
Affinity maturation in GCs of LNs• ~2M total B cells per LN• ~20K ag‐specific B cells per LN (1%)• ~200 ag‐specific CDR3 families per animal
(based on GC model of normal rodent)
Plasma or memory cell differentiation
Our analysis is focused onLN‐derived B‐cells
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Immunization & LN harvest
Immunization & LN harvest
Deep sequencing, Ranking
Deep sequencing, Ranking
Gene Assembly & Expression
Gene Assembly & Expression
Ab Characterization
Ab Characterization
Display, Select Display, Select
Sequence AnalysisSequence Analysis
Ab Characterization
Ab Characterization
Isolate Antigen Specific B‐cellsIsolate Antigen Specific B‐cells
HC Repertoire Cloning & Expression
HC Repertoire Cloning & Expression
Ab Characterization
Ab Characterization
HC Repertoire Cloning
HC Repertoire Cloning
OmniFlic antibody discovery methods
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Cloning, expression and selection in yeast
expression and identification
RT‐PCR
VH D JH
V
V2
V3
V4
V6
J or C
XhoI
cloning
fixed VL JL
````````
VH DJH
S SS S
Aga1
Aga2 HA
yeast cell
Bio
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OmniDeep™ sequence‐based discovery
Platform is a unique combination of– Antibody repertoire deep sequencing– Custom bioinformatics analysis– High‐throughput vector assembly– Recombinant expression and screening
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OmniDeep screening strategy– Primary screen: All prominent CDR3 sequence families (ELISA, affinity, functional)– Secondary screen: Complete lineages of primary hits (affinity, functional)
Primary Screen:100‐400 diverse CDR3 sequences
Guided by lineage rank analysis
SecondaryScreen:50‐300 unique sequences per lineage
Includes rare sequences in lineages of interest
hit
hit
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6 OmniFlic rats,injected with PDL1
6 OmniFlic rats,injected with PDL1
LN tissue from 2 legs X 2 tech reps of each = 4 total samples per rat
Deep sequencing and analysis done on each sample
Candidates expressed as fully human IgG with fixed light chain
Example: PDL1 antibody discovery in OmniFlic
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HCAb Repertoire Rank Analysis
1 2 3 4 5Adj only rats Adj+antigen rats
LN samples used for hybrid/yeast display
Blue= hybridoma/yeast display sequences from rat #6
Blocks of red= high freq seqs unique to one antigen rat
high freq seqs found in multiple antigen rats but not in adj-only rats
Antigen-selected samples
high freq seqs found in antigen rats and adj-only rats
1 2 3 4 5 6 7
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Selection of antibodies for primary screenHigh‐frequency sequences chosen from 2 categories– CDR3 families found in multiple rats – CDR3 families found in individual rats
234 total heavy chain sequences – 106 unique CDR3 sequence families
Expressed as fully human IgG with fixed LC in HEK cells
ELISA screen
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Primary screen: PDL1 ELISA binding & affinity
– Each row is a unique VH sequence expressed as IgG with fixed LC
– 127 of 234 total abs positive by ELISA– 62 of 106 unique CDR3 families positive by ELISA– 1.3 nM highest affinity– 2 different CDR3 sequences with ligand‐blocking
activity in cell‐based assay
Red= strong bindingBlue= negative binding
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PDL1 ligand‐blocking sequence families
Primary Screen– Identified 2 antigen‐specific
antibody families with ligand blocking activity
Secondary Screen– Identify family members with
higher affinity and fewer sequence liabilities
Primary Screen Secondary Screen
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affinity (nM) CDR seq liability1.31.32.2 met,nglyc2.7 met4.0 met5.0 nglyc6.36.87.88.08.28.4 nglyc9.0 nglyc
10.0 deam,met13.7 met,nglyc13.714.4 met from primary screen16.118.421.923.225.4 deam29.038.548.249.0 from primary screen49.553.7 nglyc79.5
PDL1 ligand‐blocking family #1
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PDL1 ligand‐blocking family #2affinity (nM) CDR seq liability
4.4 isom,cys4.9 isom from primary screen5.9 isom6.1 isom6.3 isom6.9 isom6.9 isom7.8 isom,cys8.0 isom
10.1 isom11.3 isom11.8 isom,cys12.3 isom17.5 nglyc20.6 deam,isom23.5 isom61.4 isom62.8 isom67.0 deam,isom82.0 deam,isom92.0 isom
870.0 deam,isom980.0 isom
1665.0 deam,isom1750.0 deam,isom1920.0 deam,isom2480.0 deam,isom2670.0 deam,isom2710.0 deam,isom