Information Memorandum. Opportunity for Leadership in Multiplexed Signal Detection for Molecular Diagnostics with Anapa s MELTPLEX Technology

Transcription

1 Information Memorandum Opportunity for Leadership in Multiplexed Signal Detection for Molecular Diagnostics with Anapa s MELTPLEX Technology July 2018

2 Disclaimer While the information contained in this presentation is believed to be accurate, Anapa Biotech ( Anapa ) disclaims all liability for the content of or omissions from it. The presentation includes forward looking statements and includes assumptions about future developments, operations and results. Although such statements are believed to be reasonable, there can be no assurances that such assumptions and views of the future are accurate, or that estimates and projections will be realized. No representation, warranties or other assurances will be made by Anapa concerning the anticipated performance of the company. The presentation has been prepared solely for the information of those to whom it is supplied and it should not be considered as a recommendation that any recipient of the presentation, should invest in Anapa. Prospective participants in a transaction are required to form their own opinion. 2

3 Overview Anapa develops and validates nucleic acid technologies that improve third-party PCR-based assays Anapa s strategy is to license their technologies to key molecular diagnostic providers in large markets Anapa has developed MeltPlex, a patented Taqman-type detection method, which combines fluorometric detection with meltcurve detection, increasing analyte detection by 4-5 fold. With the MeltPlex methodology, users can get faster results Achieve single tube PCR detection of 20+ targets Get more answers from a single, scarce sample eg. spinal fluid samples or biopsies Use existing Taqman target sequences Can support syndromic detection - vs single agent tests Use MeltPlex on existing instruments Adapt MeltPlex to most PCR instruments it adresses over installed instruments in the market Upgrade existing sample-to-result PCR-platforms to multiplex Save time and money with MeltPlex Save reagent costs Achieve simpler and faster lab procedures 3 3

4 Transaction Objectives Anapa Biotech ( Anapa ) is a molecular research tools company involved in developing novel, IP-protected, highly multiplexed signal detection for up to 20+ nucleic acid sequences in a single tube. Anapa is seeking expressions of interest from a partner to bring to market its first commercial product, MeltPlex. Anapa is flexible in terms of the structure of any future relationship which may include Out-licensing Acquisition of IP Co-development of current and future products Acquisition of some or all of the assets of the company 4 4

5 Resources Available to a Potential Partner Anapa provides an opportunity to gain: Market leadership with MeltPlex, a unique development for super-multiplexing of high throughput molecular diagnostics tests. Unique research tools for rapid nucleic acid detection, up to 20+ in a single tube assay. Strong global IP in this space If required, access to complete fast track assay development services by the Anapa team using the best of Anapa s MeltPlex technology 5 5

6 EXECUTIVE SUMMARY 6

7 Executive Summary Anapa Biotech ( Anapa ; is a Copenhagen, Denmark - based company with novel intellectual property and a first-in-class approach to high performance multiplexing for molecular diagnostic assays. Despite developments in conventional PCR, the complexity of multiplex Real Time PCR is still limited due to the lack of sufficient detection channels. To achieve high-end multiplexing capacity on standard Real Time PCR machines, Anapa Biotech has developed the MeltPlex technology. MeltPlex utilizes a system of fluorescently labeled Taqman-type probes allowing each to be read out by subsequent melting curve analysis. 5 or more probes can be analyzed per fluorophore channel. By utilizing melting curve readout of modified probes one for each target - the system adds an extra level of specificity to melting curve analysis as well as added multiplexing capacity. PCR reaction and melting analysis is performed without the need to re-open PCR reaction tubes Anapa develops molecular tools to enhance the performance of natural nucleic acids in PCR based assays as well as hybridization. Anapa s technology offers improvement to areas such as: Human and veterinary diagnostics Food and feed quality and safety Environmental surveillance Scientific research 7

8 MeltPlex Key Highlights Despite the developments in conventional PCR, the complexity of multiplex Real Time PCR is still limited due to the lack of sufficient detection channels. To achieve high-end multiplexing capacity on standard Real Time PCR machines, Anapa Biotech has developed the MeltPlex technology. MeltPlex utilizes a system of labelled Taqman-type probes allowing each to be read out by subsequent melting curve analysis. 5 or more probes can be analyzed per fluorophore channel. By utilizing melting curve readout of modified probes one for each target - rather than only the amplicons, the system adds an extra level of specificity to melting curve analysis. Reaction and melting analysis is performed without the need to re-open PCR reaction tubes. To test the system, Anapa designed MeltPlex probes based on previously designed TaqMan probes targeting 16 different hemorrhagic fever viruses and tested them against artificial DNA targets. 4-5 probes per fluor channel potentially higher Good success rate converting existing TaqMan probes to MeltPlex format Based on known chemistry/designs Works on most PCR cyclers Requires melting step after PCR Quantitative for single positives per channel 8

9 MeltPlex USPs Novel method allows single tube PCR detection of 20+ targets Addresses an installed base of over instruments Proof-of-concept data available In line with increased market focus on multiplex (syndromic detection vs single agents Very long lived IP and freedom to operate More answers from single (scarce) sample (Sepsis, spinal fluid, biopsies etc.) Cost efficient and simpler lab procedure lower overall costs per test Supports syndromic detection (vs. single agent tests) Option for proprietary position for key player Adaptable to most PCR instruments Upgrading of existing PCR-platforms to multiplex without altering hardware Insignificant additional reagent costs for converting existing 3-4 plex assays into one MeltPlex assay Significant value for running one test versus multiplex assays 9

10 COMPANY PROFILE 10

11 Anapa Biotech Company Profile Founded 2007 (originally named Quantibact) as a spin-out from Department of Clinical Microbiology at Hvidovre University Hospital Acquired Twisted Intercalating Nucleic Acid technology in 2007 and licensed to Eurofins and Trilink Biotechnologies for sale as a research use product Developed MeltPlex method (2015) Located in Scion DTU Science and Technology Park Main investor is Vecata Invest (> 80% owner) CEO is Soren Morgenthaler Echwald, an experienced molecular biologist 11

12 MELTPLEX WORKFLOW 12

13 MeltPlex Technology Animation We have prepared a short animated movie to explain graphically how Anapa s revolutionary MeltPlex technology works in practice. To watch this, please just click on the link below: HQAJBnw&feature=youtu.be 13

14 MeltPlex Technology STEP 1 -HYBRIDIZATION PCR primers for amplification of DNA or reverse transcribed RNA target serve as target DNA. An amplification step is performed using PCR primers and multiple target-specific MeltPlex probes labeled with fluorescent dyes. 14

15 MeltPlex Technology STEP 2 - PROBE CLEAVAGE If the specific target is present, the 5 quencher is cleaved from the probe which is now activated (fluorescent). 15

16 MeltPlex Technology STEP 3 - DETECTION A second, common, quenching probe hybridizes to the Melting Tag region and generate a melting curve with a profile specific for each target-specific probe. 16

17 MeltPlex Technology STEP 4 - READ-OUT The detection of the activated probes is done by melting curve analysis on a Real-Time PCR system. 20+ target-specific MeltPlex detection probes, labeled by multiple fluorophores and varying in Melting Tag temperature, enable specific detection of amplified product and the corresponding pathogen. 17

18 DATA ANALYSIS 18

19 Working with Ugentec Fastfinder Software

20 MELTPLEX DATA 20

21 MeltPlex Test System 28 hemmorraegic fever viruses (Pang et al, PLOS One, 2014) Artificial DNA templates w. unique primers Use pre-validated Taqman probe sequences from publication MeltPlex probes for same or complementary sequence First 21 probes made 20 working 21

22 Probe Sequence Examples HTNV: used probe sequence on reverse strand ZEBOV: used similar probe Virus strain Probe sequence Label Hantaan virus (HTNV) Original Taqman FAM-CCTGCAACAAACAGGGAYTACTTACGGCA-BHQ1 FAM/BHQ1 Tag TM Hantaan virus (HTNV) MeltPlex BHQ1-TGCCGTAAGTARTCCC[T-FAM]GTTTGTTGCAGGTACGGCCGATTAGAT/3phos... FAM/BHQ1 48 Ebola Zaire virus (ZEBOV Original Taqman Ebola Zaire virus (ZEBOV) MeltPlex FAM-CGGCAAAGAGTCATCCCAGTGTATCAAGTAA-BHQ1 FAM/BHQ1 BHQ2-CGGCAAAGAGTCA[T-Q670]CCCAGTGTATCAAGTAATACGGCCGATTAGAT/3-Sp3 Q670/BHQ

23 MeltPlex Proof of Concept Using a pre-determined set of probe design rules, 16 existing Taqman-probes designed to detect a range of hemorrhagic virus strains were converted to MeltPlex probes and combined into a single, multiplex, probe-based assay. Using 4 of the 5 channels of the Bio-Rad CFX96 instrument, we have shown proof-of-concept that the MeltPlex system can reliably detect 16 individual viral strain sequences 23

24 MeltPlex Proof of Concept QPCR with a mix of 16 MeltPlex probes and a single PCR-primer pair per well 24

25 MeltPlex read-out: Bio-Rad CFX 96 25

26 MeltPlex read-out: Bio-Rad CFX 96 26

27 Melt-Tag Distribution 46 o C 64 o C 48 o C 52 o C 54 o C 58 o C 27

28 INTELLECTUAL PROPERTY 28

29 MeltPlex IP Filings Title Country/ region Application number Filing date Publication number Publicatio n date DUAL QUENCHING ASSAY FOR MULTIPLEX DETECTION OF TARGET NUCLEIC ACIDS (priority application ) DUAL QUENCHING ASSAY FOR MULTIPLEX DETECTION OF TARGET NUCLEIC ACIDS Denmark Europe EP PA EP (A1) DUAL QUENCHING ASSAY FOR MULTIPLEX DETECTION OF TARGET NUCLEIC ACIDS Canada CA CA (A1) Dual quenching assay for multiplex detection of target nucleic acids China CN CN (A) DUAL QUENCHING ASSAY FOR MULTIPLEX DETECTION OF TARGET NUCLEIC ACIDS Japan JP JP (A) DUAL QUENCHING ASSAY FOR MULTIPLEX DETECTION OF TARGET NUCLEIC ACIDS DUAL QUENCHING ASSAY FOR MULTIPLEX DETECTION OF TARGET NUCLEIC ACIDS US 15/535,906 WO2015DK WO (A1) 29

30 Global Patent Filings for MeltPlex 30

31 IP Patents Anapa has a range of IP for the use of highly multiplexed PCR assays 31

32 MARKET OVERVIEW 3232

33 Market Dynamics for PCR in Molecular Diagnostics The quantitative PCR market is well established and predicted to gro at strong rates. Depending on the data source, the market today is estimated to be around $7USD and growing with CAGR of 9-12%: The global molecular diagnostics market size was valued at USD 6,451.5 million in 2015 and is anticipated to grow at a CAGR of 12.1% over the forecast period to Advancements in molecular diagnostics have enabled detection of various diseases that could possibly result in severe social and economic burden. The global PCR technologies market is expected to grow with a single-digit CAGR to reach $13.4 billion by The booming healthcare technology market with its demand for innovative technology for early and accurate diagnosis of life-threatening diseases, rising investments in companion diagnostics, advent of digital PCR, expiry of key-player PCR patents, and the increasing demand for PCR in genetic and molecular testing are some of the factors driving the PCR market growth. The global dpcr and qpcr market is expected to reach USD 5.31 Billion by 2022, growing at a CAGR of 8.9% during The market is served by more than 90 companies selling PCR machines which employ thermal cycling for quantitative PCR. 33

34 MeltPlex Collaboration Targets Companies without high-multiplex platforms are under pressure from market demands MeltPlex can integrate into current hardware and upgrade any platform to 20+ multiplex Top-tier companies who are players in this market include: Company/Technology Current limit New capability BD (BD Max) 4 plex 20+ plex Roche Cobas 5 plex 25+ plex Cepheid 4 plex 20+ plex ThermoFisher (QuantStudio) 4 plex 20+ plex Aries (Luminex) 5 plex 25+ plex Biorad 5 plex 25+ plex BioCartis 30-plex 100+ plex GenePOC 12-plex 50+ plex 34

35 Contact Details Anapa Biotech has appointed The Sage Group to manage its strategic partnering program. Please direct all enquiries to: US Corporate Office Wayne Pambianchi The Sage Group Inc Route 31 North #381 Clinton New Jersey USA Phone: EU Corporate Office Dr. Bill Mason Sage Healthcare Ltd. The Old Black Barns Lord s Lane, Ousden Newmarket CB8 8TX UK Phone: Mob: wtm@sagehealthcare.com Corporate Website 35