Announcement PDF ABN 16 165 160 841 29th September 2023 ASX RELEASE AACR PANCREATIC CANCER CONFERENCE PRESENTATION HIGHLIGHTS • • • Preclinical data from pancreatic cancer studies presented at premier international conference in Boston, USA. Data from these studies further support combination of narmafotinib (AMP945) with chemotherapy in pancreatic cancer. Plans are underway for a clinical trial of narmafotinib in combination with FOLFIRINOX. Melbourne, Australia: Amplia Therapeutics Limited (ASX: ATX), (“Amplia” or the “Company”) is pleased to announce that a poster, detailing a series of preclinical studies in pancreatic cancer models, was presented overnight at the AACR SPECIAL CONFERENCE IN CANCER RESEARCH: PANCREATIC CANCER meeting, being held in Boston, USA. The poster describes work conducted by our collaborators at the Garvan Institute of Medical Research, Sydney, and was presented by postdoctoral researcher Dr Kendelle Murphy. A copy of the presentation is attached to this announcement. Amplia’s CEO Dr Chris Burns commented: “We are delighted that this extensive body of research from the Garvan Institute on our best-in-class FAK inhibitor narmafotinib is being presented at such a prestigious conference. The Garvan team have done an outstanding job and this poster represents the first public disclosure of the breadth of that work demonstrating the impressive activity of the drug in these preclinical studies.” The poster shows data demonstrating that in disease-bearing mice narmafotinib (AMP945) significantly reduces pancreatic cancer associated fibrosis through inhibition of FAK activity. Further, in a number of in vivo pancreatic cancer models, narmafotinib can be utilized to improve chemotherapy response. Dr Burns continued: “This new preclinical data provides additional support for the activity of narmafotinib in combination with the two major chemotherapy regimens for pancreatic cancer. Plans are underway to start clinical trials using narmafotinib in combination with FOLFIRINOX in a similar design to the ACCENT trial.” This ASX announcement was approved and authorised for release by the Board. - End Investor Contact: Dr Chris Burns Chief Executive Officer chris@ampliatx.com Media Contact: H^CK Director, Haley Chartres haley@hck.digital +61 423 139 163 Level 17, 350 Queen Street, Melbourne VIC 3000 Email info@ampliatx.com www.ampliatx.com About Amplia Therapeutics Limited Amplia Therapeutics Limited is an Australian pharmaceutical company advancing a pipeline of Focal Adhesion Kinase (FAK) inhibitors for cancer and fibrosis. FAK is an increasingly important target in the field of cancer and Amplia has a particular development focus in fibrotic cancers such as pancreatic cancer. FAK also plays a significant role in a number of chronic diseases, such as idiopathic pulmonary fibrosis (IPF). For more information visit www.ampliatx.com and follow Amplia on Twitter (@ampliatx), Threads (@ampliatx) and LinkedIn. Pulsed priming with narmafotinib reduces fibrosis and enhances both gemcitabine/Abraxane & FOLFIRINOX response in pancreatic cancer Kendelle J Murphy*‡, Cecilia R Chambers†, Daniel A Reed†, Lily M Channon, Victoria Lee, Anna E Howell, Max Nobis, Astrid Magenau, Janett Stoehr, Brooke A Pereira, Sophie McKay, Anaiis Zaratzian, Michael Tayao, Andrew Da Silva, Australian Pancreatic Genome Initiative (APGI), Australian Pancreatic Matrix Atlas (APMA), Ruth J Lyons, Anthony J Gill, Andrew V Biankin, Jaswinder Samra, C Elizabeth Caldon, Thomas R Jeffry Evans, Lorraine Chantrill, Lisa G Horvath, Owen J Sansom, Jennifer P Morton, Tri G Phan, Yingxiao Wang, Terrie-Anne Cock, Anthony Bishop, Mark Devlin, John Lambert, Christopher J Burns, Thomas R Cox, Marina Pajic, David Herrmann*‡ and Paul Timpson*‡ Treatment naïve B.) 40 20 High Average Fiber Length 0 Mask Treatment naïve FOLFIRINOX 80 60 Gem/Abraxane Tumour Microarrays (TMAs) FOLFIRINOX Second Harmonic Generation (SHG) **** **** E.) 40 20 0 -20 Gem/Abraxane Treatment naïve Treatment naïve FOLFIRINOX Gem/Abraxane FOLFIRINOX Low pTyr-397-FAK High pTyr-397-FAK F.) Low Average Fiber Length High Average Fiber Length G.) Low Average Fiber Length Matrix Contraction/Fibrotic Remodeling High Average Fiber Length H.) 400 +/- narmafotinib 75 300 ** * 200 *** *** 100 50 0 *** 25 0 *** I.) 500 Area (mm2) Probability of Survival (%) 226 Patients 100 +/Neoadjuvant Chemotherapy Mask 60 -20 Gem/Abraxane SHG Coverage (%) Here, we provide preclinical data showing that the ATP-competitive, orally bioavailable small molecule, narmafotinib; a highly potent and selective inhibitor of FAK, with desirable potency, selectivity, and pharmacokinetics (PK) in healthy human volunteers, which can be utilized to improve chemotherapy response in pancreatic cancer. 80 C.) 100 Vehicle J.) 0 1000 2000 3000 4000 5000 Vehicle 5 nM 10 nM 20 nM 50 nM 100 nM 5 10 20 50 narmafotinib [nM] K. Contraction: Day 12 Days Probability of Survival (%) +/Neoadjuvant Chemotherapy Picrosirius Red Coverage (%) 100 Low Average Fiber Length Picrosirius Red and TWOMBLI A.) **** **** Picrosirius Red pFAK immunohistochemistry • Pancreatic ductal adenocarcinoma (PDAC) is a highly fibrotic and aggressive malignancy with a poor 5-year survival rate of 12%. • The standard-of-care for these patients is largely limited to one of two systemic chemotherapy regimens: Gemcitabine in combination with Abraxane (nab-paclitaxel) or FOLFIRINOX regimen consisting of oxaliplatin, irinotecan, leucovorin and 5-fluorouracil. • PDAC progression is accompanied by a fibrotic response in the stroma, involving the elevated deposition and remodeling of extracellular matrix (ECM), promoting tissue stiffness and driving tumor progression as well as poor chemotherapeutic response. • Focal Adhesion Kinase (FAK) is a non-receptor tyrosine kinase that modulates bidirectional interactions between cancer cells and their adjacent ECM via integrin-mediated signaling. • We and others have recently shown that FAK inhibition can reduce fibrosis, supporting the clinical assessment of FAK inhibition for cancer treatment. Average Fiber Length D.) ECM deposition and organization correlates with PDAC patient surivival and outcomes in combination with FAK signature. Maximum SHG Intensity [AU] Fibrotic Targeting in PDAC Introduction 150 100 ns ** 50 ** *** 0 Vehicle 100 nM ** 5 10 20 50 narmafotinib [nM] 100 nM Second Harmonic Generation (SHG) Vehicle 5 nM 10 nM 20 nM 50 nM 100 nM Low Average Fiber Length Low p-Tyr-397-FAK 100 High Average Fiber Length High p-Tyr-397-FAK 75 50 25 * 0 0 1000 2000 3000 4000 5000 Days (A.) Quantification, and representative images of Picrosirius Red (B.) SHGimaging of tissues from PDAC patients (C.) Schematic representation of patient TMA establishment and staining with Picrosirius Red. (D.) Kaplan-Meier analysis of human PDAC survival based on average collagen fiber length (E.) Schematic representation of patient TMA staining with pTyr-397-FAK and Picrosirius Red. (F.) Kaplan-Meier analysis of human PDAC survival (G.) Schematic of 3D organotypic contraction assay. (H.) Quantification of matrix contraction at day 12 and (I.) SHG maximum intensity of contracted matrices at day 12. (J.) Representative images of matrix contraction (scale, 1 cm), and (K.) SHG maximum intensity (scale, 100 µm), at day 12. Personalised Medicine E.) ns 0.6 Vehicle narmafotinib Late narmafotinib Early narmafotinib Early narmafotinib Priming: 10 mg/kg Oral Administation Subcutaneous injection 1x106 KPC-FAK Cells Timeline am Day 6 Day 7 Day 8 Day 9 3.5 narmafotinib 3.5 Tumour Growth MP 840 nm MP 840 nm Inactivation narmafotinib FRET Y397 SH2 FRET P 1.5 1.5 Inactive G.) am pm Active Vehicle Day 0 pm am pm FLIM-FRET Y397 ECFP Fluorescence Lifetime (ns) H.) J.) * 20 Days post Injection 30 Vehicle narmafotinib 2.3 2.2 2.1 2.0 Vehicle narmafotinib Day 0 Timeline am pm am pm Collagen (SHG) Vehicle Gem/Abraxane narmafotinib G2/M narmafotinib Gem/Abraxane Day 6 Day 7 Day 8 Day 9 Day 11 Day 10 L.) Vehicle Gem/Abraxane narmafotinib narmafotinib Gem/Abraxane G1 G0 M.) Cleaved Caspase-3: Apoptosis Vehicle Gem/Abraxane 50 *** ** 4 100 Time post treatment (days) Vehicle/Saline 100 75 0 25 50 75 100 Days 125 150 175 200 Subcutaneous Injection Time line Tumor Establishment **** 4 Vehicle *** 3 2 1 0 narmafotinib Gem/Abraxane - + - + * + + narmafotinib H.) 80 **** 30 20 + + C. 20 + - + + + narmafotinib + FOLFIRINOX Cleaved Caspase-3 10 * 15 10 5 narmafotinib Gem/Abraxane - + - + + + (A.) Schematic of 3D organotypic invasion assay; fibroblast-driven matrix contraction, cancer cell seeding and invasion into the matrix under a chemotactic gradient. (B.) Quantification and (C.) representative images of KPC cell invasion into 3D organotypic matrices (scale bar, 100 µm). (D-E.) Kaplan-Meier analysis of survival in mice with KPC subcutaneous tumors treated with (D.) Priming or (E.) Chronic vehicle (orange), narmafotinib (blue), gemcitabine/Abraxane alone (light green) or narmafotinib priming prior to gemcitabine/Abraxane (dark green) (F). Timeline of subcutaneous KPC-FAK xenograft establishment, treatment and intravital imaging in live tumors surgically exposed via skin flap to monitor FAK activity using the (G.) Lyn-FAK-FRET biosensor. (H.) Representative images and quantification of ECFP fluorescence lifetime (nanoseconds, scale bar 50 μm) in mice treated with vehicle or narmafotinib (10mg/kg). (I.) Representative images of SHG maximum intensity projections (scale bar, 50 μm) and quantification of maximum SHG signal intensity (J.) Timeline of subcutaneous KPC-FUCCI xenograft establishment, treatment schedule and intravital imaging in live tumors surgically exposed via skin flap to monitor cell cycle distribution using the FUCCI cell cycle reporter. (K.) Representative FUCCI images and quantification of cell cycle distribution in tumors treated with vehicle or narmafotinib followed by saline or gemcitabine/Abraxane (scale bar, 100 µm). (L.) Representative maximum intensity SHG images and quantification of peak SHG signal intensity (scale bar, 100 µm). Representative images and quantification as a percentage of positive cells for (M.) cleaved caspase-3 and (N.) Ki67 (scale bar, 50 µm) stained tumors. 0 10 20 30 40 50 Time post treatment (days) 20 FOLFIRINOX 5 100 0 FOLFIRINOX narmafotinib + FOLFIRINOX Vehicle/Saline ** **** Vehicle/FOLFIRINOX narma/FOLFIRINOX ** narma/Saline 75 50 0 -100 5000 10000 Placebo 25 50 100 narmafotinib (AMP945) [mg] 125 15000 -50 -100 -50 narmafotinib (AMP945) Plasma Exposure (AUC, ng.h/mL) (A.) Chemical structure of narmafotinib. (B.) pTyr-397-FAK levels in skin punch biopsies from healthy volunteers at 2-4 hours post narmafotinib administration. (C.) Linear relationship between change in FAK activity from baseline and AUC0-inf following dosing of narmafotinib. AUC0-T: Area under the plasma concentration-time curve from the time of dosage administration over the dosage interval. -Patients will recieve one-week priming dosage of narmafotinib (once daily oral capsule). Gemcitabine and Abraxane will then be given according to standard-of care chemotherapy and patients given pulsed dosing of narmafotinib for four days prior to weekly chemotherapy. 50 Narmafotinib Priming: 7 Day Loading 25 0 FOLFIRINOX narmafotinib FOLFIRINOX r2 = 0.37 p = 0.005 0 -Pulsed dosing regimen of narmafotinib in combination with gemcitabine and Abraxane in first-line chemotherapy for patients with unresectable or metastatic pancreatic cancer. TKCC 10lo I.) ** 25 25 0 100 50 ACCENT Trial (NCT05355298) 40 15 narmafotinib Gem/Abraxane *** 50 I.) 0 + Day 12 60 0 + - Day 9 75 ns FOLFIRINOX narmafotinib FOLFIRINOX - Day 8 Ki67 10 0 - am am am pm pm pm Day 1 Day 2 Day 3 Day 4 AM: Oxaliplatin PM: Irinotecan Repeated pulsed treatment cycle every 12 days until endpoint Ki67: Proliferation Vehicle Gem/Abraxane narmafotinib Priming: 10 mg/kg Oral Administation Vehicle narmafotinib FOLFIRINOX narmafotinib/ FOLFIRINOX 100 FOLFIRINOX 40 narmafotinib Gem/Abraxane N.) G.) Calcium Leucovorin 5-Fluorouracil 100 25 narmafotinib + Gem/Abraxane *** B. Narmafotinib 50 0 Gem/Abraxane * **** narma/Gem/Abraxane * narma/Saline A. Vehicle/Gem/Abraxane 2 F.) * 25 Ki67 Positive Cells (%) narmafotinib narmafotinib Gem/Abraxane Cleaved Caspase-3 Positive Cells (%) Vehicle * 50 SHG Intensity [AU] S 75 narmafotinib Gem/Abraxane Second Harmonic Generation: Collagen Vehicle M G2 narmafotinib + Gem/Abraxane Cleaved Caspase-3 6 0 Day 12 Tumour Growth Skin flap 80 FOLFIRINOX G1/G0 100 IP Injection Gem/Abraxane am Gem/Abraxane 0 Gem/Abraxane narmafotinib Gem/Abraxane Vehicle narmafotinib 60 5 8 16 40 10 24 20 20 TKCC 10lo D.) * 0 E.) 15 0 28 0 ** Gem/Abraxane narmafotinib Gem/Abraxane SHG 25 Ki67 20 40 Intravital Imaging pm ** narmafotinib Gem/Abraxane FUCCI Cell Cycle Reporter Vehicle Subcutaneous injection 1 x 106 KPC Cells C.) Gem/Abraxane 50 G1/S K.) narmafotinib Priming: 10 mg/kg Oral Administation * * 25 SH2 FAK Activation narmafotinib 10 Day 12 Repeated pulsed treatment cycle every 12 days until endpoint Vehicle I.) 2.5 2.4 Chronic 0 Day 10 Day 7 Time line Tumour Establishment 50 Skin flap Intravital Imaging 40 75 0 F.) 30 ** pFAK + + + - 20 Days post Injection am am am pm pm pm Day 4 Day 1 Day 2 Day 3 75 Probability of Survival + - 100 10 IP Injection Gem/Abraxane IP Injection Gem/Abraxane Vehicle/Saline narmafotinib/Saline Vehicle/GemAbraxane narmafotinib/GemAbraxane 100 Probability of Survival narmafotinib Early narmafotinib Late 0 narmafotinib Priming 10 mg/kg Oral Administation Subcutaneous Injection Ki67 Positive Cells (%) 0.8 * Probability of Survival ** B.) 25 0 ns 1.0 50 • Narmafotinib (Amplia Therapeutics Ltd) was discovered through a structure-guided drug discovery program. • ATP competitive small molecule inhibitor of FAK with a potency for FAK (IC50 2.2nM) and is selective over the closely related kinase Pyk2 (IC50 550nM). • Narmafotinib displays high selectivity across a panel of 468 kinases, along with favourable pharmacokinetics in murine species. • In mice, orally dosed narmafotinib possesses a half-life of ~3 hours and oral bioavailability of 47%. • A Phase I randomized double-blind, placebo-controlled study in 56 healthy volunteers was undertaken (ACTRN12620000894998) to investigate safety, tolerability, Pharmacokinetic and Pharmacodynamic effects of orally dosed drug. • The single ascending dose study showed that narmafotinib could be detected in the plasma over 24 hours following administration at various dosages as low as 15 mg up to 125 mg. • Dose and exposure-response in skin punch biopsies following a single treatment with narmafotinib showed drug target engagement through reduction of pTyr-397-FAK levels, with a clear dose demonstrated by a significant linear relationship between change in FAK activity from baseline. • No serious or severe treatment emergent adverse events (TEAE)s, nor any TEAEs leading to study withdrawal were observed and the majority of TEAEs reported were mild. pFAK C.) 1.2 *** narmafotinib Gem/Abraxane 27% Gemcitabine and Abraxane A.) Probability of Survival Collagen Matrix Invasion Normalised Invasive Index B.) Invasion Air-Liquid Interface Gem/Abraxane 75 Probability of Survival Metal Grid * * narmafotinib Ki67 Positive Cells (%) KPC Cell +/- narmafotinib (late) Vehicle Cleaved Caspase-3 Positive Cells (%) Invasion Priming 100 Cleaved Caspase-3 Positive Cells (%) +/- narmafotinib (early) Seed Cancer Cells Fibroblast % Cell Cycle Phase Vehicle/ narmafotinib washout Matrix Contraction/Fibrotic Remodeling D.) 14 Days SHG Intensity [AU] 3 Days 12 Days Probability of Survival A.) Narmafotinib priming prior to gemcitabine/Abraxane or FOLFIRINOX chemotherapy significantly extends survival in long-term orthotopic studies using patient-derived models. Change from baseline (%) Narmafotinib priming reduces FAK activity and fibrosis whilst improving the response of KPC cells to gemcitabine/Abraxane Clinical Translation: ACCENT Trial Change from baseline (%) Priming 0 25 50 75 100 Days 125 150 175 (A.) Schematic of treatment schedule and timeline for TKCC10lo subcutaneous survival study and (B.) Kaplan-Meier analysis of survival in mice with TKCC10lo subcutaneous tumors Representative images and quantification as a percentage of positive cells for (C.) Ki67 and (D.) cleaved caspase-3 stained tumors after 3 cycles of treatment (E.) Kaplan-Meier analysis of survival in mice with TKCC10lo orthotopic tumors (F.) Schematic of treatment schedule and timeline for TKCC10lo subcutaneous survival study and (G.) Kaplan-Meier analysis of survival in mice with TKCC10lo subcutaneous tumors. Representative images and quantification as a percentage of positive cells for (H.) Ki67 and (I.) cleaved caspase-3 stained tumors after two cycles of treatment with FOLFIRINOX (J.) Kaplan-Meier analysis of survival in mice with TKCC10lo orthotopic tumors. Chemotherapy Suttons Group Narmafotinib Priming Narmafotinib Priming Narmafotinib Priming Chemotherapy Chemotherapy Leonard Ainsworth ACRF:INCITe Centre