Anaptysbio, Inc. · NASDAQ:ANAB
Anaptysbio — Festival of Biologics 2020
16 slides from Anaptysbio's conference presentation, Mar 2, 2020
Slide 1: Discovery of a PD-1 Checkpoint Agonist Antibody for Autoimmune/Inflammatory Disease
Discovery of a PD-1 Checkpoint Agonist Antibody for Autoimmune/Inflammatory Disease Marilyn Kehry, Ph.D. Vice President, Cell and Functional Biology March 2nd 2020
Slide 2: PD-1 is an Inhibitory Checkpoint Molecule that Down-modulates T Cell Responses
PD-1 is an Inhibitory Checkpoint Molecule that Down-modulates T Cell Responses • Immune checkpoint receptor-ligand interactions are essential for T cell APC down-regulating immune responses and maintaining self-tolerance CD226 PVR PVRL2 Co-Stimulatory OX40 OX40L • Functional antagonist antibodies to
Slide 3: PD-1 Agonist Antibody Discovery
PD-1 Agonist Antibody Discovery ANB030 is a Humanized IgG1,κ Hybridoma-derived PD-1 Agonist Antibody sPD-1-Fc Hybridoma PD-1+ cells PD-1+ cell binders + Fusion screened for PD-1 ligand competition Immunization PD-L1 non- blocking hybridomas Optimization for Tm, affinity, developability Purified anti
Slide 4: Optimization of ANB030 VH Sequence for Stability and Affinity
Optimization of ANB030 VH Sequence for Stability and Affinity Saturating Mutagenesis at 4 VH Positions Humanized VH CDRH1 CDRH2 CDRH3 Humanized CDRL1 CDRL2 CDRL3 VL Affinity by KinExA KD KD ANB030 Variant Human Cynomolgus Fab Tm Antibody PD-1 PD-1 Mouse chimeric 4.3 nM n.d. n.d. CDR-grafted 0.19 nM
Slide 5: ANB030 Binding to PD-1 Does Not Compete with PD-L1 Binding
ANB030 Binding to PD-1 Does Not Compete with PD-L1 Binding Ligand non-blocking is desirable to avoid potential antagonist activity PD-1 agonist discovery for autoimmune/inflammatory disease 5
Slide 6: Model of the Epitope Bound by PD-1 Agonist Antibody ANB030
Model of the Epitope Bound by PD-1 Agonist Antibody ANB030 • Regions on PD-1 bound by sans loop ANB030 were identified by hydrogen-deuterium exchange (pink, blue) and PD-1 surface point mutations (red) • Both datasets generated a consistent model of the ANB030 epitope PD-1 PD-1 mutations that elimin
Slide 7: ANB030 in Solution Potently Inhibits Whole Blood Tetanus Toxoid Recall Response
ANB030 in Solution Potently Inhibits Whole Blood Tetanus Toxoid Recall Response IL-2 Whole blood Tetanus Toxoid C o n t r o ls PD-1 Agonist ANB030 Anti-PD-1 1500 1 5 0 0 1500 1 5 0 0 IgG4 Isotype IgG1 Isotype Anti-CD3ε & Ab IFNγ( p(pg/ml) 4d g /m l) 1000 coat (pg/ml) 1 0 0 0 IF N (p g /m l) 1000 1
Slide 8: ANB030 Functional Agonist Activity in Whole Blood is Dependent on FcγR Engagement
ANB030 Functional Agonist Activity in Whole Blood is Dependent on FcγR Engagement IL-2 Whole blood Wild Type IgG1 IgG2 Isotype Ig G 2 Is o ty p e A N B 0 3 0 Tetanus Toxoid W T I gANB030 G 1 A N B 0 3 0 ANB030 1500 1 5 0 0 1500 1 5 0 0 Anti-PD-1 Anti-CD3ε & Ab IgG2 Isotype 4d p g /m l) coat p g /m l
Slide 9: Alopecia Areata Is An Immune-Mediated Disease
Alopecia Areata Is An Immune-Mediated Disease Alopecia areata is an immune-mediated form of hair loss resulting from breakdown of immune privilege that is driven by keratinocyte and melanocyte antigen-specific T cells producing IFNγ Healthy Hair Follicle Alopecia areata Hair Follicle • Low MHC class
Slide 10: Inhibition of IFNγ Production by ANB030 in Human Alopecia Areata PBMCs
Inhibition of IFNγ Production by ANB030 in Human Alopecia Areata PBMCs Stimulated with Keratinocyte Peptide Antigens Alopecia areata is an immune-mediated form of hair loss resulting from breakdown of immune privilege that is driven by keratinocyte and melanocyte antigen-specific T cells producing I
Slide 11: PD-1 Signaling in T Cells
PD-1 Signaling in T Cells Bardhan K, Anagnostou T and Boussiotis VA (2016) Front. Immunol. 7:550. PD-1 agonist discovery for autoimmune/inflammatory disease 11
Slide 12: ANB030 Induces SHP2 but not SHP1 Recruitment to PD-1 after Activation of Jurkat PD-1 Cells
ANB030 Induces SHP2 but not SHP1 Recruitment to PD-1 after Activation of Jurkat PD-1 Cells +αCD3 +αCD3 PD-1 Jurkat WB + kDa αCD3 PD-1 αPD-1 49 beads PD-1 IP SHP2 SHP2 62 SHP2 SHP1 62 2 minutes 10 minutes Lyse cells, add anti-PD-1 beads to isotype samples for PD-1 immunoprecipitation ANB030 had no ef
Slide 13: NSG/Hu-PBMC Graft vs. Host Disease Model
NSG/Hu-PBMC Graft vs. Host Disease Model Endpoints: • Weight loss (-20%) • Death • GvHD scores Weight and GvHD assessment 3X weekly FACS FACS FACS NSG mice n=12/group d.7 d.14 d.21 d.28 d.35 d.42 Day 0: Irradiation 0.9 x 107 hu PBMC i.v. mAb dosing 24-hr-post PBMC Groups: 1. Isotype control IgG1 2.
Slide 14: PD-1 Agonist Antibody ANB030 is Efficacious in an Acute Xenogeneic Graft vs. Host Disease Model
PD-1 Agonist Antibody ANB030 is Efficacious in an Acute Xenogeneic Graft vs. Host Disease Model G v H D K a p la n -M e ie r P lo t L a s t d o s in g d a y 1 0 0 Ig G 1 Is o ty p e P e r c e n t s u r v iv a l A N B 0 3 0 Treatment Median 5 0 Survival * p = 0 .0 2 * Isotype 20 days ANB030 39.5 days
Slide 15: Summary: PD-1 Agonist Discovery for Autoimmune/Inflammatory Disease
Summary: PD-1 Agonist Discovery for Autoimmune/Inflammatory Disease • A functional agonist anti-PD-1 antibody that down-regulates antigen-specific immune responses and lacks antagonist activity has been discovered and optimized • ANB030 is a humanized IgG1/κ anti-PD-1 agonist antibody that is non-bl
Slide 16: Acknowledgments
Acknowledgments • Cell Biology: Allison Rooks, Jack Manorek, Natasha Del Cid, Eric Hare, Nina Schommer • Protein Sciences: Larry Altobell, Robert Morse, Greg Gold • Molecular Biology & Protein Expression: Geoff Tomlinson, Brandon Dolan • Stephen Parmley, V.P. Molecular Biology & Protein Sciences • M