Slide 1: Expanding new target space inside cells with biologics using Phylomer peptides
For personal use only Expanding new target space inside cells with biologics using Phylomer peptides Phylogica: Bio, June 2016
Slide 2: Disclaimer
Disclaimer The purpose of the presentation is to provide an update of the business of Phylogica Limited (ASX:PYC) For personal use only [‘Phylogica’]. These slides have been prepared as a presentation aid only and the information they contain may require further explanation and/or clarification. Acc
Slide 3: About Phylogica
About Phylogica § Owns unique proprietary class of peptide therapeutics (Phylomers) For personal use only § Most structurally diverse source of peptides available for drug discovery § Validated platform for discovery and superior delivery of peptide drugs against intracellular targets § Advancing on
Slide 4: Majority of drug targets (80%) are inside cells
For personal use only Majority of drug targets (80%) are inside cells Distribution of drug targets Small Molecules Cell Membrane Outside Cells Inside Cells Biologics (20%) (80%) (10%) Market for biologics drugs $220b $0 § PYC addressing unmet need to deliver biologic drugs inside cells! 4
Slide 5: Technology Introduction
For personal use only Technology Introduction
Slide 6: Phylomers: peptides from parts of biodiverse proteins
Phylomers: peptides from parts of biodiverse proteins § Encoded by >35 biodiverse microbial genomes For personal use only § Most diverse source of natural secondary/tertiary structures - evolved to bind protein interfaces and for functional activity § Delivers very high functional rates for drug dis
Slide 7: Phylogica: Unique capabilities for intracellular discovery and delivery of biologics drugs
Phylogica: Unique capabilities for intracellular discovery and delivery of biologics drugs Delivery 1 For personal use only Unique Targets Discovery 2 3 Potential to expand the druggable intracellular landscape by >10-fold 9
Slide 8: The Problem: Biologics cargos are trapped within endosomes
The Problem: Biologics cargos are trapped within endosomes For personal use only Drug Drug Drug contacts cell Budding membrane endosome Contact triggers formation of endosomes Cell Membrane § Conventional CPPs normally only active at Endosome concentrations of >10mg/Kg, limiting feasible clinical ap
Slide 9: The Solution: The Endosomal Escape Trap
For personal use only The Solution: The Endosomal Escape Trap RBD Phylomer Avitag Biotin Capture of escaped Phage Trapped Phylomer SA T7 Phage Library RBD Phylomer Avitag RBD Phylomer Avitag RBD Phylomer Avitag Release and Capture (Streptavidin Beads) Escaped Phylomer BirA § PYCs endosome escape tra
Slide 10: Split-GFP complementation assay to validate endosomal escape
Split-GFP complementation assay to validate endosomal escape of penetrating Phylomer/cargo fusions For personal use only FACS CPP RBD GFP S11 + Cell viability GFP 1-10 GFP S11 !! !! !! GFP signal !! !! !! !! !! Live Cell Microscopy !! Uptake Complementation § Only biologics delivered to cytoplasm ca
Slide 11: Phylomer is best-in-class for delivery of protein into cells
% complemented cells Phylomer is best-in-class for delivery of protein into cells (HCC-827) 100 Protein Conc. 20µM +ve) Signal (%) For personal use only 80 10µM 160 fold % Green cells (SplitGFP 5µM 37 fold 2.5µM 60 1.25µM GFP Fluorescence 0.667µM 40 20 In vitro complementation assay 0 84 22 35 9 n P
Slide 12: Delivering biologics cargoes into cells using Phylomer FPPs
For personal use only Delivering biologics cargoes into cells using Phylomer FPPs Peptides Toxins FPP Scaffolds Oligonucleotides
Slide 13: Delivering the pro-apoptotic peptide PAP
Delivering the pro-apoptotic peptide PAP § Mitochondriotoxic PAP domain (KLAKLAK)2 induces mitochondrial apoptosis1 For personal use only - Decreased proliferation of triple negative breast cancer cell line (MDA-MB-468) PHYLOMER FPP – GAS– (KLAKLAK)2 CARGO MDA468 48h Viability - 10µM MDA468 48h Viab
Slide 14: Delivering the pro-apoptotic peptide PAP
Delivering the pro-apoptotic peptide PAP § Potent functional activity of a pro-apoptotic peptide1 For personal use only § FPP-PAP more potent than TAT-PAP, especially at lower concentrations 100 MDA468: Total apoptosis % Total Apoptosis (AnnV+ve) 80 PAP 60 TAT-PAP FPP-PAP 40 20 0 2.0 2.5 3.0 3.5 4.0
Slide 15: Developing best in class Myc inhibitors using Phylomer FPP delivery
For personal use only Developing best in class Myc inhibitors using Phylomer FPP delivery
Slide 16: MYC: A classic cancer target
MYC: A classic cancer target § MYC: Amplified/overexpressed in most human cancers Proliferation For personal use only Cell Growth MYC Metabolism Apoptosis § MYC: non redundant role in cancer – associated with addiction § Considered undruggable – lacks active sites. Role for PPIs § OmoMyc*: 90 aa pep
Slide 17: Delivering Omomyc using Phylogica's delivery solution
Delivering Omomyc using Phylogica’s delivery solution For personal use only § Linked a Phylomer cell penetrating peptide (FPP PYJ02) to Omomyc Cell Penetrating Phylomer PYJ02 Omomyc § Tested for activity in a triple-negative breast cancer cell line resistant to conventional drugs § Potency measured
Slide 18: FPP-OmoMYC kills triple negative breast cancer cells
FPP-OmoMYC kills triple negative breast cancer cells use only MDA468 Omyc 4.5uM MDA468 Omyc 0.6uM 150 150 Omyc Omyc PYJ02_Omyc PYJ02_Omyc % Viability % Viability 100 100 *** *** For personal 50 50 *** **** 0 0 48h 72h 48h 72h § Potent activity against drug resistant (triple negative) breast cancer c
Slide 19: Activity of FPP-Omomyc in a range of Myc-driven cancers
Activity of FPP-Omomyc in a range of Myc-driven cancers For personal use only 1746-Omyc Cell line Disease Cell line characteristics (48h: IC50 ) T47D Breast cancer p53 mut(hetero), Myc+++ 6.6 Breast cancer p53 mut(hetero), Myc+, triple -ve MDA-MB-468 1.7 SUM159 Breast cancer Basal, Triple –ve 1.1 Br
Slide 20: RNA-seq profiling of T11 triple negative breast cancer cells following treatment
RNA-seq profiling of T11 triple negative breast cancer cells following treatment Down-regulated Genes Up-regulated Genes For personal use only Log2FC>1.5 P<0.05) FPP-Omomyc Omomyc PBS (3/6 hrs) FPP (3/6 hrs) (3/6 hrs) (3/6 hrs) § Treatment of T11 breast cancer cells with FPP-Omomyc for 3 or 6 hrs §
Slide 21: FPP-OmoMYC targets known MYC pathway genes
FPP-OmoMYC targets known MYC pathway genes § Differentially regulated genes include known Myc targets For personal use only – RNA-seq profiling of T11 cells treated with FPP-Omomyc for 3 or 6 hrs § Identifies both up- and down-regulated Myc-associated genes 3 hours 6 hours fold-change fold-change fo
Slide 22: FPP-Omomyc inhibits growth of tumours, even after treatment
FPP-Omomyc inhibits growth of tumours, even after treatment T11 Syngeneic triple negative breast cancer graft For personal use only 1200 1000 Tumour Volume (mm3) 800 PBS (vehicle) 600 Omomyc Intratumoural Injections 1746 FPP 400 1746-Omomyc FPP 200 0 0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 Day p
Slide 23: FPP-mediated delivery of Omomyc in breast cancer allograft model
FPP-mediated delivery of Omomyc in breast cancer allograft model Tumour Cell Death Proliferation Immunity H&E 1.5x H&E 10x H&E 20x Ki67 20x PDL1 20x For personal use only PBS (Vehicle) FPP Omomyc Omomyc FPP § Increased cell death, decreased proliferation & PDL1 expression in treated tumours § Potent
Slide 24: EµMyc Lymphoma Model for validation of FPP-MYC hits
EµMyc Lymphoma Model for validation of FPP-MYC hits For personal use only Inject mice IV daily with Phylomer FPP-Omomyc fusion EuMyc cells/mouse (in 100uL PBS) At each time point: D0 D3 D5 D7 D9 - 2 mice per group are culled and weighed - Spleens (and weigh) D7:and Extract cells from spleen bone mar
Slide 25: Phylogica's oncology pipeline
For personal use only Phylogica’s oncology pipeline
Slide 26: Phylogica's Intracellular Oncology Pipeline
Phylogica’s Intracellular Oncology Pipeline § Phylomer screens against 4 validated and clinically relevant oncology targets For personal use only § Validated hits already exceed potency of gold standard therapies Functional Program Target Indications Discovery Preclinical In vitro ValidationIn vivo
Slide 27: Proprietary iMYCs show equivalent potency to OmoMYC
Proprietary iMYCs show equivalent potency to OmoMYC use only § IC50 in T47D cells at 48 hrs 48h IC50s 200 For personal 150 FPP_Omomyc ICIC50 50 = 4.5µM = 7.315 % Viability FPP_iMyc1 ICIC50 50 = 3.5µM = 5.176 100 ICIC50 50 = >35µM Omomyc = 36.21 50 0 -3 -2 -1 0 1 2 Conc. (uM) 45
Slide 28: FPP-iMyc conjugates: Serum stability (0-24h)
For personal use only FPP-iMyc conjugates: Serum stability (0-24h) SA/Ni-CHL AlphaLISA: Stability in 100% FCS at 37°C SA/Ni-CHL AlphaLISA: Stability in 100% FCS at 37°C 0-3 hour serum incubation 0 -24 hour serum incubation 200000 200000 Stability (AL units) FPP-iMyc2 Stability (AL units) 150000 FPP-
Slide 29: High affinity primary 'iMyc' Phylomers against Myc
High affinity primary ‘iMyc’ Phylomers against Myc § Solution affinities determined by microscale thermophoresis For personal use only § Primary hits before any sequence maturation exceed that of Omomyc and native interaction with Max transcription factor iMyc-0113 iMyc-0114 Omomyc Max Affinity (Kd)
Slide 30: Modular approaches to extending serum half-life
Modular approaches to extending serum half-life § Assessing different modules to extend extend serum half life For personal use only – Albumin binding domains – PASylation § Addition of PK modules has minimal effect on in vitro potency § Pilot study confirms ABD can extend serum half-life Albumin Bi
Slide 31: Summary
Summary § Generating ‘first-in-class’ therapies against challenging intracellular For personal use only target classes, intractable to small molecule therapies § Confirmed ability to expand druggable intracellular target landscape § Advancing proprietary oncology pipeline towards clinical developmen
Slide 32: Phylogica is open to discovery alliances with Pharma
Phylogica is open to discovery alliances with Pharma § Phylogica’s business model is open to alliances at any stage: For personal use only – Discovery alliances – Alliances around internal programs § Open to multiple licensing structures: – Option to license – License for particular cargoes or targe
Slide 33: Contact Details
Dr. Richard Hopkins Contact Details Dr. Paul Watt Chief Executive Officer Chief Scientific Officer Tel: +61 8 9489 7777 Tel: +61 8 9489 7777 Fax: +61 8 9489 7700 For personal use only Fax: +61 8 9489 7700 Mobile: +61 405 656 868 Mobile: +61 421 550 213 richardh@phylogica.com paulw@phylogica.com www.