Nurix Therapeutics, Inc. · NASDAQ:NRIX

Nurix Therapeutics — Dana Farber TPD Webinar (February 2023)

28 slides from Nurix Therapeutics's conference presentation, Feb 2, 2023

  1. Slide 1: Discovery and Optimization of CBL-B Inhibitors

    Discovery and Optimization of CBL-B Inhibitors Dana Farber TPD Webinar Feb 2, 2023

  2. Slide 2: Important Notice and Disclaimers

    Important Notice and Disclaimers This presentation contains statements that relate to future events and expectations and as such constitute forward-looking statements within the meaning of the Private Securities Litigation Reform Act of 1995. When or if used in this presentation, the words “anticipa

  3. Slide 3: Nurix's DELigase Protein Modulation Discovery Platform

    Nurix’s DELigase Protein Modulation Discovery Platform DEL Discovery Rational and Direct-to-Cell Scaled Screening for Empirical Chemistry Biology Capabilities in vivo exposure M e an U n b o u n d D ru g F re e D r u g C o n c e n tra tio n (u M ) C o n c e n tr a tio n in P la s m a 1 0.1 0.01 0.00

  4. Slide 4: Nurix Is Advancing Four Wholly Owned Clinical Programs with a Deep Pipeline

    Nurix Is Advancing Four Wholly Owned Clinical Programs with a Deep Pipeline of Proprietary and Partnered Novel Targets TPD targeted protein degradation; TPE targeted protein elevation; TPM targeted protein modulation 4

  5. Slide 5: CBL-B is a Modulator of Immune Cell Activation

    CBL-B is a Modulator of Immune Cell Activation • CBL-B is an E3 ubiquitin ligase highly expressed in cells of the immune system CBL-B inhibition • CBL-B regulates T, B, and NK cell activation IL-2 production Proliferation Central memory phenotype • Blocking CBL-B removes a brake on the immune system

  6. Slide 6: CBL-B Is a Modulator of Immune Cell Activation

    CBL-B Is a Modulator of Immune Cell Activation Inactivation or deletion of CBL-B results in hyperactive T cells and inhibition of tumor growth IL-2 secretion in KO and ligase inactive T cells ex vivo Ligase-inactive cbl-b knock-in mice inhibit tumor growth (TC-1 syngeneic model) Het Inactive ligase

  7. Slide 7: Inactive CBL-B Is Autoinhibited

    Inactive CBL-B Is Autoinhibited RING • When Y363 of CBL-B is not phosphorylated, the helix of the LHR domain packs against the TKB domain • Incapable of binding Ub-E2 LHR • Phosphorylation of Y363 requires dissociation of LHR- RING from TKB Y363 TKB Dou, et. al., Nature Structural & Molecular Biolog

  8. Slide 8: Active CBL-B Binds Ub-loaded E2 Ligases

    Active CBL-B Binds Ub-loaded E2 Ligases “Closed-state” “Open-state” (inactive) (active, binds E2-Ub) RING UbcH5b Y363 RING LHR Phosphorylation pY363 LHR TKB TKB RING TKB Ub LHR CBL-B TKB RING unstructured 36 427 982 8

  9. Slide 9: Multiple Lead-Finding Approaches Afforded CBL-B Binders

    Multiple Lead-Finding Approaches Afforded CBL-B Binders N-Avi Biochemical Ub HTS DEL Fragment RING Screening TKB Lib size 300K 1X109 1600 TKB SRC kinase RING P # of 1 2 1 Series E2~Ub Binding Assay Hit 28 mM 2.4 mM 1800 mM Affinity DEL Ligand Cube Plot Hit mwt 338 537 211 Hit LE 0.27 0.22 0.33 Affin

  10. Slide 10: CBL-B HTS Triage Revealed a Singleton Hit

    CBL-B HTS Triage Revealed a Singleton Hit 104 hits from Closed State HTS Next: Tested from purchased powders • CBL-B Phosphorylation FRET assay ~90% confirmed from powders • E2~Ub Binding FRET assay Next: Hits re-tested from purified compounds • Src Counter Screen FRET assay < 15 hits repeated from

  11. Slide 11: HTS Reveals a Singleton Hit

    HTS Reveals a Singleton Hit E2 binding FRET, Orthogonal Src Counter Screen FRET FRET Artifact Assay 5 0 1 0 0 50 Inactive Inactive A c t iv it y A c t iv it y NRX-1 % A c t iv it y 5 0 (racemic) 0 0 % IC50: 28 µM mwt = 338 0 .0 1 0 .1 1 10 100 0 .0 1 0 .1 1 1 0 1 0 0 % 0 0 .0 0 1 0 .0 1 0 .1 1 1 0 1

  12. Slide 12: NRX-3 Is a Specific Inhibitor of CBL-B

    NRX-3 Is a Specific Inhibitor of CBL-B 1 0 0 SPR A c t iv it y 5 0 % NRX-2 0 Chiral SFC 0 .0 0 1 0 .0 1 m M 0 .1 C o m p o u n d 1 1 0 1 0 0 NRX-1 HTS Screening hit KD: 16 µM 1 0 0 A c t iv it y NRX-3 5 0 Resolved Screening hit % E2-Ub: IC50 = 12 mM 0 mwt = 338; LE = 0.29 0 .0 0 1 0 .0 1 m M 0 .1 C

  13. Slide 13: NRX-3 Is an Intramolecular Glue

    NRX-3 Is an Intramolecular Glue Substrate protein E2 E2 Y363 p Y363 1. Kinase pY363 RING TKB TKB 2. E2/substrate TKB Closed State Opened State Phosphorylation locks INACTIVE CBL-B in the ACTIVE Conformation Immune Response 13

  14. Slide 14: NRX-3 Is an Intramolecular Glue

    NRX-3 Is an Intramolecular Glue Substrate protein E2 E2 Y363 p NRX-03 RING Y363 RING 1. Kinase pY363 TKB TKB TKB 2. E2/substrate TKB NRX-3 acts as an Closed State Opened State Phosphorylation locks intramolecular glue INACTIVE CBL-B in the ACTIVE forcing CBL-B in its Conformation folded INACTIVE sta

  15. Slide 15: Crystal Structure Confirms Binding Mode as Intramolecular Glue

    Crystal Structure Confirms Binding Mode as Intramolecular Glue 2.0 Å NRX-3 binds to closed-state CBL-B and prevents phosphorylation 15

  16. Slide 16: Early SAR: Focus on Affinity and Properties

    Early SAR: Focus on Affinity and Properties NRX-3 NRX-4 NRX-5 NRX-6 E2-Ub: IC50 (mM) 12 0.23 0.092 0.088 Ligand Efficiency 0.29 0.33 0.36 0.37 Cellular Substrate Ub IC50 (mM) 7 3 1.7 Microsomes h/m Clint 20/360 -/500 30/73 (mL/min/kg) Plasma stability m/r T1/2 (min) - 140/- 280/- Papp MDCK (MDR1) A→

  17. Slide 17: Early SAR: Focus on Affinity and Properties

    Early SAR: Focus on Affinity and Properties NRX-6 NRX-7 NRX-8 E2-Ub: IC50 (mM) 0.088 0.038 0.021 Ligand Efficiency 0.37 0.37 0.36 Cellular Substrate Ub IC50 (mM) 1.7 0.78 0.79 Microsomes h/m Clint (mL/min/kg) 30/73 -/67 7/26 Plasma stability m/r T1/2 (min) 280/- >1000/163 >1000/>1000 Papp MDCK (MDR1

  18. Slide 18: Complex SAR for Rat Plasma Stability

    Complex SAR for Rat Plasma Stability The SAR for rat plasma stability was not predictable by chemists First observed with low recovery in PPB assays 18

  19. Slide 19: Machine Learning Model for Rat Plasma Stability

    Machine Learning Model for Rat Plasma Stability To assist with lead optimization, models were built based on the 104 experimental plasma stability data points available at the time Despite the low volume of data, both regression and classification models demonstrated high predictive power and provid

  20. Slide 20: NRX-8 Is a Specific Inhibitor of CBL-B

    NRX-8 Is a Specific Inhibitor of CBL-B CEREP Panel, <40% activity at 10 mM (N = 52) CBL-B SPR characterization Time (s) 40% NRX-8 displays clean 1:1 binding stoichiometry with CBL-B and is clean in off-target screening 20

  21. Slide 21: NRX-8 Maintains Original Hit Binding Mode

    NRX-8 Maintains Original Hit Binding Mode 21

  22. Slide 22: NRX-8 Inhibits Substrate Ub and Stimulates IL-2 Induction

    NRX-8 Inhibits Substrate Ub and Stimulates IL-2 Induction Human T cell assay – IL-2 production Substrate Ubiquitylation – BT20 cell line % Ubiquitylation Activity 0 -50 -100 -4 -3 -2 -1 0 1 2 3 2.5X Log uM NRX-8 IL-2 (2.5X over baseline 80 nM response) Cellular Ubiquitylation of substrate 850 nM (BT

  23. Slide 23: Pharmacologic Inhibition of CBL-B Recapitulates Anti-Tumor Effects of Genetic Model

    Pharmacologic Inhibition of CBL-B Recapitulates Anti-Tumor Effects of Genetic Model of Ligase Inhibition Ligase-inactive cbl-b knock-in mice inhibit CT26 Syngeneic Model tumor growth in TC1 Syngeneic Model 2500 Vehicle Tumor volume mm3 45 mpk BID 2000 90 mpk BID (MeanSEM) 1500 180 mpk BID 1000 500

  24. Slide 24: Over 10,000-fold Enzymatic Potency Improvement Achieved

    Over 10,000-fold Enzymatic Potency Improvement Achieved While Improving Molecular Properties 2017 HTS hit HTS hit, single enantiomer 2018 2019 ~ 10,000x potency improvement Enzymatic potency (uM) Clinical candidate range LipE 24

  25. Slide 25: Single-Agent NX-1607 Induces Antitumor Response in Multiple Models

    Single-Agent NX-1607 Induces Antitumor Response in Multiple Models NX-1607 NX-1607 NX-1607 Reduced Tumor Volume Prolonged Survival Reduced Tumor Volume Colorectal Triple-Negative Breast B Cell Lymphoma 100 Vehicle NX-1607 75 % Survival p=<0.0001 50 25 0 0 25 50 75 100 125 150 Days post implant NX-16

  26. Slide 26: NX-1607 and Anti-PD-1 Synergize To Enhance Anti-tumor Effects

    NX-1607 and Anti-PD-1 Synergize To Enhance Anti-tumor Effects and Survival of Mice in Multiple Tumor Models Colorectal (CT26) Colorectal (MC38) Triple-Negative Breast (4T1) Day 28 4T1 Lung Metastases p<0.01 # Metastatic Colonies 15000 p<0.01 10000 5000 200 100 0 Vehicle NX-1607 anti-PD-1 NX-1607+ant

  27. Slide 27: Summary

    Summary ▪ CBL-B regulates T, B, and NK cell activation ▪ Multiple screening approaches afforded validated binders to CBL-B ▪ Plasma instability may be an under-appreciated liability for amide-containing compounds ▪ Pharmacological inhibition of CBL-B recapitulates the anti-tumor effects of the genet

  28. Slide 28: Thank You

    ”) nurix Thank You

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