Repotrectinib/AMG510, 30 mg/kg vs AMG510, 30 mg/kg, p= 0.0138

Repotrectinib Increases KRASG12C Inhibitor Effectiveness Via Simultaneous Inhibition of SRC, FAK, and JAK2

Figure 1. Repotrectinib Kinase Selectivity

• KRAS regulates a complex signaling network with multiple downstream pathwaysthat promote survival, proliferation, and cytokine secretion in cancer cells1-7

• Resistance via feedback reactivation, bypass signaling, and cytokine-mediatedtumor immunity limit the efficacy of KRASG12C inhibitors (AMG510, MRTX849) assingle agents1,2

• SRC, FAK, and JAK2 play important roles in the KRAS signaling network andoperate in a compensatory manner to affect both tumor cell intrinsic and extrinsicprocesses3-9

• Simultaneous inhibition of SRC/FAK/JAK2 by repotrectinib has the potential toimprove the effectiveness of KRASG12C inhibitors

Brion W. Murray, Wei Deng, Dayong Zhai, Nathan V. Lee, Laura RodonTurning Point Therapeutics, Inc., San Diego, CA 92121

INTRODUCTION

EVALUATION OF REPOTRECTINIB IN BIOCHEMICAL ASSAYS

Abstract # 147

• Repotrectinib (Figure 1A) is a selective kinase inhibitor currently in a registrationalclinical trial for ROS1+ and NTRK+ cancer patients10

• Repotrectinib potently inhibits SRC/FAK/JAK2 in biochemical assays (Figure 1B‑C)

Presented at: EORTC-NCI-AACR (ENA) Symposium on Molecular Targets and Cancer Therapeutics, 24 – 25 Oct 2020. Corresponding Contact — Jim Mazzola: jim.mazzola@tptherapeutics.com

1. Canon J, et al., 2019, Nature 575(7781), 217-223

2. Hallin J, et al., 2020, Cancer Discov 10(1), 54-71

3. Lee HJ, et al., 2014, Cancer Cell 26(2), 207-21

4. Sen B, et al., 2009, Cancer Res 69(5), 1958-65

5. Wilson C, et al., 2014, Oncotarget 5(17), 7328-41

6. Murakami Y, et al., 2017, Oncotarget 8(41), 70736-70751

7. Zhou Q, et al., 2017, J Pharmacol Exp Ther 363(3), 428-443

8. Kitajima S, et al., 2016, Semin Cell Dev Biol 58,127-35

9. Ji et al., 2019, Proc Natl Acad Sci USA 116(19), 9453 – 9462

10. Drilon et al., 2018, Cancer Discov 8(10), 1227 - 1236

11. Ianevski A, et al., 2017, Bioinformatics 33(15), 2413-2415

CONCLUSIONS

REFERENCES & DISCLOSURES

• Repotrectinib inhibits SRC/FAK/JAK2 in in vitro and in vivo models• SRC, FAK, STAT3 and AKT phosphorylation is suppressed by repotrectinib in in vitro

and in vivo models

• In preclinical models, repotrectinib synergizes with AMG510 and inhibits KRASG12C

tumor cell proliferation, suppresses receptor tyrosine kinase upregulation induced by AMG510, and reduces KRASG12C tumor cell cytokine release. Similar results were obtained with other KRASG12C inhibitors.

• Repotrectinib significantly enhances efficacy of AMG510 and shows a marked survival benefit in a KRASG12C xenograft model

• Additional preclinical studies are on-going with AMG510 and other KRASG12C

inhibitors• Potential combination regimens for repotrectinib are under evaluation for clinical

development

EVALUATION OF REPOTRECTINIB/AMG510 IN CELL-BASED ASSAYS

• Repotrectinib/AMG510 combination potently inhibits activation of SRC, FAK, STAT3,AKT, and ERK in H358 KRASG12C NSCLC cells (Figure 2)

• Repotrectinib/AMG510 combination suppresses more KRAS signaling nodes thanAMG510 combinations with dasatinib, defactinib, or ruxolitinib in H2122 KRASG12C

NSCLC cells and therefore modulates a broader signaling network (Figure 3)

• Repotrectinib/AMG510 combination leads to a decrease in H358 KRASG12C NSCLCcell viability (Figure 4A) compared to single-agent treatment. Similar results wereobserved in other KRASG12C cell lines (H2122, H23, H1792, H1373)

• Repotrectinib/AMG510 combination is synergistic in H2122 KRASG12C NSCLC cells(Figure 4B)

• Repotrectinib suppresses an adaptive receptor tyrosine kinase feedback loopinduced by AMG510 in KRASG12C NSCLC cells (Figure 5)

• Repotrectinib inhibits cytokine secretion and enhances AMG510 effects in KRASG12C

NSCLC cells (Figure 6)

Figure 2: Repotrectinib/AMG510 Inhibition in H358 KRASG12C NSCLC Cells

EVALUATION OF REPOTRECTINIB/AMG510 IN CELL-BASED ASSAYS

Figure 3: Repotrectinib/AMG510 Combination vs AMG510 Combinations with Dasatinib, Defactinib or Ruxolitinib Inhibition in H2122 KRASG12C NSCLC Cells

Figure 4: REPOTRECTINIB/AMG510 EFFECTS ON KRASG12C CELL VIABILITY

EVALUATION OF REPOTRECTINIB/AMG510 IN CELL-BASED ASSAYS

Figure 5: Repotrectinib/AMG510 Combination Effect on Receptor Tyrosine Kinase Activation in KRASG12C NSCLC Cells

Figure 6: Cytokine Secretion and Enhanced AMG510 Effects in KRASG12C Cells

EVALUATION OF REPOTRECTINIB/AMG510 IN IN VIVO TUMOR MODELS

C.B.A.

*Proxy reagents purchased from commercial sources

*Proxy reagents purchased from commercial sources

AMG510 Repotrectinib BLISS Synergy Score

41 nM 1000 nM 21

123 nM 1000 nM 16

A. B.

Cytokine array in H2122 KRASG12C media

(IL‑6 highlighted)

RNA expression of cytokines in H2122 KRASG12C NSCLC cells

This study was sponsored by Turning Point Therapeutics, Inc.

BWM, WD, DZ, NVL, LR:  Turning Point Therapeutics – employment/shareholder.

Figure 7: Evaluation of SRC/FAK/JAK2 Signaling in In Vivo Assays

EVALUATION OF REPOTRECTINIB/AMG510 IN IN VIVO TUMOR MODELS

Target IC50 (nM) at 10 µM ATP

ROS1 0.07TRKC 0.21TRKB 0.3TRKA 0.53ALK 1.0JAK2 1.0

SRC & SRC family kinases

1.1–5.3

TXK 3.2ARK5 4.5DDR1 5.7

FAK 7.0

4 hours+ - - -- - + +- + - +

48 hours+ - - -- - + +- + - +

Control

AMG510* (0.1 µM)

Repotrectinib (1 µM )

pERK-T202/Y204

ERKpAKT-S473

AKTpSTAT3-Y705

STAT3pFAK-Y397

FAKpSRC-Y416

SRCActin

- + - + - + - + - +

- - + + - - - - - -

- - - - + + - - - -

- - - - - - + + - -

- - - - - - - - + +

AMG510* (KRASi, 0.1 µM)Repotrectinib

(SRCi/FAKi/JAK2i, 1 µM)Dasatinib* (SRCi, 0.1 µM)

Defactinib* (FAKi, 1 µM)

Ruxolitinib* (JAK2i, 1 µM)

pFAK-Y397

FAK

pSTAT3-Y705

STAT3

pAKT-S473

AKT

pERK-T202/Y204

ERK

Actin

24 hours

H2122H358

Figure 8: Survival Effect of Repotrectinib/AMG510 in H2122 NSCLC KRASG12C Xenograft Tumor Model

• At clinically relevant repotrectinib exposure, repotrectinib/AMG510 combinationpotently inhibits SRC, FAK, STAT3, ERK, and AKT phosphorylation in a H358 KRASG12C

cell-derived xenograft tumor model (Figure 7)

• Repotrectinib significantly enhances survival for AMG510 treatments in an H2122xenograft model (Figure 8)

IL‑6 ELISA in KRASG12C Media

Quantitation of cytokine secretion in KRASG12C

cell media