touchlight

APPLICATIONS OF DBDNA™
DR. LISA CAPRONI
DISCLAIMER
This document is a presenta1on which has been prepared by, and is the sole property of, Touchlight Gene1cs Limited (the "Company"). The informa1on in this document is subject to upda1ng, comple1on, revision, further verifica1on and amendment. This document does not purport to contain all the informa1on that a prospec1ve investor may require. This printed presenta1on is incomplete without reference to the oral presenta1on, discussion and any related wriDen materials that supplement it. Prospec1ve collaborators of the Company should conduct their own independent inves1ga1on and analysis of the informa1on contained in this document and they are advised to seek their own professional advice on the legal, financial and taxa1on consequence of entering into any such collabora1on. No reliance may be placed for any purpose whatsoever on the informa1on contained in this document or on its completeness. No undertaking, representa1on or warranty, express or implied, is given by the Company, any member of its Group or any of their respec1ve current or proposed directors, officers, partners, employees, secondees, agents or advisers or any other person as to the accuracy or completeness of the informa1on or as to the opinions contained in this document and no liability is accepted for any such informa1on or opinions. This document and its contents are confiden1al and neither it nor any copy of it may be distributed, re-­‐distributed, published or reproduced or otherwise made available in whole or in part or disclosed by recipients to any other person and, in par1cular, it may not be distributed to persons without the prior consent of the Company. This document includes statements that are, or may be deemed to be, "forward-­‐looking statements". These forward-­‐looking statements can be iden1fied by the use of forward-­‐looking terminology, including the terms "believes", "envisages", "es1mates", "an1cipates", "projects", "expects", "intends", "may", "will", "could", "seeks" or "should" or, in each case, their nega1ve or other varia1ons or comparable terminology, or by discussions of strategy, plans, objec1ves, goals, future events or inten1ons. These forward-­‐looking statements include maDers that are not historical facts and speak only as of the date of this document. They appear in a number of places throughout this document and include statements regarding the Company and the directors' current inten1ons, beliefs or expecta1ons concerning, amongst other things, investment strategy, financing strategy, performance, results of opera1ons, financial condi1on, liquidity, prospects, growth, strategies and the industry in which the Company operates. By accep1ng a copy of this document, you agree to be bound by the foregoing limita1ons and, in par1cular, will be taken to have represented, warranted and undertaken that you have read and agree to comply with the contents of this no1ce including without limita1on the obliga1on to keep this presenta1on and its contents confiden1al. INTRODUCTION TO TOUCHLIGHT
DNA PLAYS A KEY ROLE THROUGHOUT THE BIOTECH INDUSTRY
Current uses of DNA in the Biotechnology industry Biologics DNA vaccines Gene therapy Cell therapy RNA Industrial biomaterials Agriculture !   Transient and stable transfec1on of microbial and mammalian cell lines for the produc1on of therapeu1c proteins and mAbs !   In vivo expression of biological products from injected DNA !   Direct injec1on of DNA constructs to elicit an immune response for either therapeu1c or prophylac1c effect in animals or humans !   Direct injec1on of DNA designed to express a given protein !   Large-­‐scale transfec1on of pDNA into AAV and Len1viral vectors for gene therapy !   pDNA transfected via viral vectors to produce modified CAR T cells !
pDNA for stem cell reprogramming !   In vitro transcrip1on from a DNA template to produce mRNA (useful in nucleic acid vaccines but also in gene edi1ng systems) !   In vivo transcrip1on of RNAi from a DNA template (miRNA, siRNA) !   Synthe1c metabolic pathways to generate novel biomaterials !   DNA bioelectronics !   Gene1c modifica1on of crop plants PDNA APPLICATIONS
PDNA IS CRITICAL FOR DNA VACCINES, GENE & CELL THERAPY
pDNA Gene therapy Cell therapy DNA vaccines !  AAV vector transduc1on !  CAR T cells !  DNA administered directly !  Len1viral vector transduc1on !  TCR cells !  Virus-­‐free gene therapy !  CRISPR !  ~100 products in clinical trials !  ~250 products in clinical trials !  ~50 products in clinical trials !  1 product (Glybera) marketed in Europe !  Mul1ple FDA breakthrough designa1ons !  Inovio’s VGX-­‐3100 enrolling Phase III GMP manufacture of plasmid DNA is primarily for preclinical and clinical-­‐stage products COPYRIGHT TOUCHLIGHT GENETICS LTD. 2015. CONFIDENTIAL
PDNA MANUFACTURING
COMMERCIALISATION OF GENE THERAPIES WILL CAUSE PDNA BACKLOG
Clinical trials Preclinical Animals Early stage Commercialisa1on Late stage Today’s pDNA capacity can meet this demand, but a backlog of mul;ple months exists Orphan drug CHF drug Significant pDNA capacity increases required COPYRIGHT TOUCHLIGHT GENETICS LTD. 2015. CONFIDENTIAL
PDNA MANUFACTURING
PLASMID DNA MANUFACTURE CAN BE INEFFICIENT AND PROBLEMATIC
Plasmid DNA manufacture CLONING !   Bacterial plasmids can be problema1c to purify at scale ESTABLISH CELL LINE - 
Amplifica1on and cell lysis >6 weeks - 
Large fermenta1on volumes required - 
Purifica1on of pDNA from chromosomal DNA and bacterial endotoxin is troublesome !   GMP pDNA presents regulatory ques1ons PLASMID AMPLIFICATION - 
Contain bacterial sequences - 
Contain an1bio1c resistance genes !   pDNA manufacture is expensive and a risk to COGS of cell and gene therapies PLASMID PURIFICATION !   Can Touchlight’s technology ease the boDleneck and provide func1onal advantages in terms of efficacy? FINAL PLASMID PRODUCT COPYRIGHT TOUCHLIGHT GENETICS LTD. 2015. CONFIDENTIAL
DBDNA™ FROM TOUCHLIGHT
GENETICS
TOUCHLIGHT’S DBDNA™ IS PRODUCED IN A TWO ENZYME PROCESS
Protelomerase binding
sequence
START
Φ29 DNA polymerase,
dNTPs and primer
Denature
Rolling circle amplification
of DNA template
Strand displacement
DNA “plasmid” template
Denature
Protelomerase
DNA concatamer
Cuts and ligates DNA
dbDNA™ can be used
as a starting template
by the DNA polymerase
dbDNA™
COPYRIGHT TOUCHLIGHT GENETICS LTD. 2015. CONFIDENTIAL
DBDNA™ FROM TOUCHLIGHT
GENETICS
DBDNA™ CASSETTES HAVE MANY BENEFITS OVER PLASMID
dbDNA™ expression casseBes Func@onal benefits Regulatory benefits !   Sequence length can exceed plasmid (>13kb) !   No bacterial sequences !   Highly stable due to closed ends !   Can be generated in fully bacteria-­‐free system !   Very low error rate !   No an1bio1c resistance gene !   Sequences with complex secondary structure possible !   ‘Toxic” genes no problem COPYRIGHT TOUCHLIGHT GENETICS LTD. 2015. CONFIDENTIAL
DBDNA™ FROM TOUCHLIGHT
GENETICS
DBDNA™ CAN MEET FUTURE DEMAND
pDNA ReacAon volume dbDNA™ >10,000L fermenta1on <500L reac1on PurificaAon procedure Mul1ple large-­‐scale ion exchange and size exclusion chromatography steps Simple precipita1on procedure Infrastructure required Large-­‐scale (>2,000L) agitated bioreactors; Large-­‐scale chromatography equipment Minimal >6 weeks <2 weeks Time to pure product Total pure product 1kg COPYRIGHT TOUCHLIGHT GENETICS LTD. 2015. CONFIDENTIAL
DBDNA™ FROM TOUCHLIGHT
GENETICS
CAN DBDNA™ REPLACE PLASMIDS IN VIRUS MANUFACTURE?
Proposed dbDNA™ system Plasmid system (generic) 10
Producer cells Producer cells Pseudoviral par1cles Pseudoviral par1cles !   BoDleneck in GMP plasmid manufacture !   Scalable produc1on of DNA casseDes !   Bacterial sequences and an1bio1c resistance genes can be incorporated into final product !   No bacterial sequences or an1bio1c resistance COPYRIGHT TOUCHLIGHT GENETICS LTD. 2015. CONFIDENTIAL
DBDNA™ FROM TOUCHLIGHT
GENETICS
DBDNA™ CASSETTES EXPRESS FUNCTIONAL PROTEIN IN VIVO
dbDNA™ influenza vaccine (ScoD et. al., 2015) tel
L
CMV IE
PR8 HA
SV40 late
Poly A
tel
R
PBS (not challenged)
Naïve
dbDNA™ pDNA
!   Codon and RNA-­‐op1mised PR8 HA gene encoded on a dbDNA™ !   25ug dose given to mice at day 0, 21 and 42 !   Mice given lethal PR8 influenza viral challenge at day 91 !   Bodyweight and survival post-­‐challenge measured, in addi1on to immunological measures !   dbDNA™ at least equivalent to pDNA vaccines COPYRIGHT TOUCHLIGHT GENETICS LTD. 2015. CONFIDENTIAL
PROBLEMS AND SOLUTIONS
!  The demand for high quality and GMP grade DNA is currently accelera1ng !  Current and near future infrastructure able to support early clinical material or orphan drug requirements but not beyond !  Our DNA planorm has advantages in scale of manufacture -­‐ small footprint, mul1 product manufacturing suites !  The lack of an1bio1c resistance, CpG islands and origins of replica1on in our DNA circumvents the growing concern with the use of these sequences in therapeu1cs !  The lack of the backbone sequences results in a smaller construct which has been shown to aid in the transfec1on efficiency