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Jan 4, 2017 (OTCQX: DYAI) Producing Biologics with C1
Jan 4, 2017(OTCQX: DYAI)
Producing Biologics with C1
Safe Harbor Regarding Forward-Looking Statements
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Certain statements contained in this presentation are forward-lookingstatements within the meaning of the federal securities laws. Theseforward-looking statements involve risks, uncertainties and other factorsthat could cause Dyadic’s actual results, performance or achievements tobe materially different from any future results, performance orachievements expressed or implied by such forward-looking statements.Any forward-looking statements speak only as of the date of thispresentation and, except as required by law, Dyadic expressly disclaimsany intent or obligation to update or revise any forward-looking statementsto reflect actual results, any changes in expectations or any change inevents. Factors that could cause results to differ materially are discussedin Dyadic’s publicly available filings, including information set forth underthe caption “Risk Factors” in our September 30, 2016 Quarterly Reportfiled with the OTC Markets on November 10, 2016 and our December 31,2015 Annual Report filed with OTC Markets on March 29, 2016. New risksand uncertainties arise from time to time, and it is impossible for us topredict these events or how they may affect us.
C1 – the most productive fungal expression system for biologics
3
A safe and reliable protein production platform, C1-derived vaccine showed no adverse clinical effects in mice5 Safety &
Reliability
Produces batches of proteins that are significantly purer than traditional production methods 12 Purity
Cuts preparation and production time in half compared to CHO3 Time Saving
Achieves much higher yields than traditional production systems in CHO, E. coli, S. cerevisiae, P. pastoris1 Yield
Grows under broader temperature & pH ranges and is easily scalable compared to CHO4 Robustness
C1 can minimize CapEx investments, production costs of biologics and overcome the limitations of traditional production systems
1 including CHO, E. coli, S. cerevisiae, P. pastoris
Dyadic Overview
Revolutionary protein expression technology “C1”: based on Myceliopthora thermophila fungus
Technology covered by over 20 patent families
Listed on the stock exchange (OTCQX: DYAI), liquidity of > 60m USD (1)
Experienced management & board– 20+ Years of Experience with Fungal Production Systems– 20+ Years in Pharmaceuticals
20+ Years of Commercial Enzyme Production
Platform optimized 2009 – 2015 Hyper productive strain developed with
unparalleled purity: >100 g/l with ~80% purity
Production approved as safe by FDA Produced in up to 500,000l tanks
Biopharmaceuticals
Strategic focus since 2016 Powerful molecular toolbox enables
production of complex proteins Application proven successful:
mAbsVaccinesNon-
GlycosylatedProteins
Dyadic has demonstrated the power of C1 for the production of biologics and is now looking to establish partnerships with biopharmaceutical companies
(1) As of September 30, 2016, including ~ $7.4 million of restricted cash held in escrow until July 2017 from the DuPont Transaction.
4
0
5
10
15
20
25
30
35
40
Per.C6 CHO C1 ComplexBiologics Yield
3
10
30
9
15
< 60
0
10
20
30
40
50
60
S. cerevisiae P. pastoris C1Industrial/Simple
Protein Yield
C1 produces more protein
Simple/Non-Glycosylated Proteins Complex Proteins, e.g. mAbs
Yiel
d in
g/l
>3 times higher yield 2 to 10 times
higher yield
Sources: 1 Boehringer Ingelheim, BioXcellence production: www.bioxcellence.com. & Shane Cox Gad (2007) Handbook of Pharmaceutical Biotechnology. Wiley Interscience, New Jersey. 2 Non-GMP conditions, non purified. 3 Susan Gotensparre (2007) Crucell. InPharmaTechnologist.com. 4 Non-GMP conditions, not purified, expected based on small scale production experience
High cell density attainable More protein produced Protein is secreted Both for small and large scale production Codon optimization established For heterologous proteins of both bacterial and mammalian origin
High productivity of C1 proven
5
1
2
34
2 3
20
KeyHighest yield claimedRealistic estimate
1
40
15
20
C1 delivers higher purity of protein and is highly robust Pu
rity
C1 White Strain 2.0 2
C1 1st
Generation2E. Coli 1
C1 delivers Higher levels of the target protein Significantly higher purity
C1 maintains high productivity Under a wider temperature than CHO Under a wider pH range than CHO At scales ranging from laboratory shake flasks to
20,000l tanks and above
40ºC37ºC-32ºC25ºC
pH range71 14
5 9C1 CHO
Rob
ustn
ess
CHO 2
1 After 2 Purification Steps, 2 No purification steps 3 Optimal range 32 - 37ºC. Source: Sellick, C. et al (2009) Optimizing CHO Cell Culture Conditions. Genetic Engineering and Biotechnology News Tutorial.
6C
HO
3
45ºC-
25ºC
C1
Reproduction rate of cell 2x higher than for CHO Protein production rate at least 1.5 fold Higher purity of protein achieved may decrease recovery time
C1 enables shorter production cycles in comparison to CHO
1
1
2
2
3
3
0 1 2 3 4 5
CHO
C1
Duration of Steps in Production
*Note: Protein Recovery may be faster due to higher purity of C1 production
Week 1 Week 2 Week 3 Week 4
Batch Cycle time is reduced by >50% in comparison to CHO, freeing up capacity
Production time reduced by >14 days
1: Biomass Expansion 2: Protein Production 3: Protein Recovery*
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C1’s unique morphology enables non-viscous fungal production
The low viscosity allows C1 to be used in established microbial production facilities,
requiring no additional CapEx investment
8
Visc
osity
(cP
)
Protein Yield
500
400
300
200
100
100
80
60
40
20
Pro
tein
(g/l)
Viscosity
Standard Fungal Line C1
Low Viscosity, High Yield
Filamentous fungi face challenges for their use in production due to high viscosity
C1 exhibits a unique morphology resulting in low viscosity
*
C1 enables 50 - 85% reduction in manufacturing costs
Expected OpEx savings of 60 - 85% vis-à-vis CHOPossibility of decreasing CapEx investments by 80 %
4
12
0
5
10
15
Column1
Decrease direct labor cost by >80% Decrease indirect labor cost by >80% Decrease consumables cost by >90% Decrease capital charge by 60-70%2
Same extraction and purification costs 3
g/10
00 U
S $
1 Depreciation cost of facility over 10 years included, savings increase with increased production need 2 Depending on production requirements, investment into 10,000 l tanks can be reduced to 1 – 2,000l tanks, 3 Cost savings likely higher due to higher purity of proteins from C14 Based on production needs of ~800kg,
Output ~tripled
Cost Efficiency of C1 vs. CHO(Exemplary ~20kg mAb Production1)
Lower CapEx Investment Smaller production facilities faster to build
Additional benefits
CapEx Investment of C1 vs. CHO(Exemplary large scale mAb Production4)
0
100
200
300
400
500
CHO C1
Milli
on U
S $
CHO C1
CapEx reduced by 80%
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OpEx upstream savings Four 10,000l tanks
Three 2,000l tanks
C1 is easy to engineer to achieved desired protein profile1
Dyadic has experience with each of the molecular tools necessary to optimize the strain for high productivity and functionality for the targeted protein class
Genetic manipulation
Computational biology
Man9 G0 G2F
Changing the cellular
regulatory circuit
✔
Libraries of efficient strong
promoters
✔
Libraries of TF and signal
peptides and / or carrier proteins
✔
Libraries of protease
deletion strains
✔
Glycoengineering to form mammalian-like
glycan structures in progress
✔
1. First proof of concept studies have been successful for these tools in Trichoderma. 10
Gene 1Pr Carrier
Trichoderma Reesi Fungal SystemProduction yields with different target proteins by Trichoderma reesei
Trichoderma Source: PEGS Boston, 2016: (Next - Generation Biotherapeutic Production System: The Filamentous Fungus Trichoderma Reesi)
Antibodies produced in Δ7 strain, IFN in Δ9 strain, and IGF1 in Δ13 deletion strain as fusion with CBHI carrier
However, this is far from the maximal theoretical output of 29 g/L for MAb01 based upon carrier expression level
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Trichoderma Reesi Fungal System (Cont.)
IgG production by Trichoderma reesei (TR)
Trichoderma Source: PEGS Boston, 2016: (Next - Generation Biotherapeutic Production System: The Filamentous Fungus Trichoderma Reesi)
Protease deletion strains together with fermentation optimization work improved the IgG
Antibody production levels up to 7.1 g/l
The secretion carrier CBHI is produced in the fermentations at levels up to 38 g/l. This theoretically equates
to potential antibody expression levels of approximately 29 g/l
12
6
18
28
34
41
49
52
6056 56
58
5.9
13.1
22.4
36.331.4
38.5
33.3 3235.4
5.8 6.2 6.7 6.9 7.1 6.74.7
17
27.623.9
29.225.3 24.3
26.9
0
10
20
30
40
50
60
70
0 1 2 3 4 5 6 7 8 9 10 11 12 13
Prot
ein
(g/L
)
Day in culture
TR - Total Protein
CBHI
TR - MAB01
TR - Theoretical MAB01
6
18
28
34
41
49
52
6056 56
58
5.9
13.1
22.4
36.3
31.4
38.5
33.3 3235.4
17
27.623.9
29.225.3 24.3
26.9
5.8 6.2 6.7 6.9 7.1 6.74.7
0
10
20
30
40
50
60
70
0 1 2 3 4 5 6 7 8 9 10 11 12 13
Prot
ein
(g/L
)
Day in culture
TR - Total Protein
CBHI
TR - Theoretical MAB01
TR - MAB01
Higher potential production of IgG with C1
(1) Expression of heterologous fungal proteinTrichoderma Source: PEGS Boston, 2016: (Next - Generation Biotherapeutic Production System: The Filamentous Fungus Trichoderma Reesi)
Total C1 cell protein expression reached a level of >100 g/l.
The production level is almost 2 fold higher than the total protein production of T. reesei.
The production phase starts earlier already after 1 day and can proceed to 5 – 7 days.
Therefore, the potential of reaching much higher antibody productivity than 6.9 g/l is promising with C1 after
5-7 days of optimized fermentation process.
13
1.7
10.1
41
66
82
95
105
6
18
2834
41 49
52
6056 56 58
10
22
34
55.2
47.8
58.4
50.648.6
53.8
5.9
13.1
22.4
36.3 31.438.5
33.3 3235.4
511
17
27.623.9
29.225.3 24.3 26.9
11.612.4 13.4 13.8 14.2 13.4 9.4
5.8 6.2 6.7 6.9 7.1 6.74.7
0
20
40
60
80
100
120
0 1 2 3 4 5 6 7 8 9 10 11 12 13
Prot
ein
(g/L
)
Day in culture
C1 - Total Protein
TR - Total Protein
C1 - Theoretical
CBHI
TR - Theoretical MAB01
C1 - Expected
TR - MAB01
C1 for vaccine production – lower costs & higher efficacy
Easy scale up, lower production costs due to higher yield (in comparison to CHO/yeast/ E. coli)
C1 produced antigen generated an equal, or better, immune response in mice than the industry standard antigen
International collaboration ongoing in vaccine development
Key Advantages
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The C1 technology platform:
A leap in technology that shows the potential to change the way in which both Human and Animal Health Biopharmaceutical companies bring their biologic vaccines and drugs to market faster, in greater volumes, at lower cost, and with newer beneficial properties.
C1 has the potential to address several of the challenges facing the Diabetes market
1. High demand Insulin production needs are expected to exceed 16 tons by 2025. Current Insulin
production methods will not suffice to meet demand.1
1. Lower margins Published insulin CoGS lie at about 61 USD per gram, leaving a profit margin of about
35%.2
1. Payer pressures US payer pressures are driving down profit margins even further.
C1 for production of non-glycosylated proteins*
Based on production yields alone, cost savings of > 50% may be achieved.
C1 exhibits higher production flexibility than S. cerevisiae or E. coli
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* With no to minimal post-translational modifications needed to functionality
Sources: 1. Baeshen et al. Cell factories for insulin production. Microbiol Cell Factories 2014. 13:141. 2 Harrison et all, Bioseparations Science and Engineering. Oxford University Press 2nd Edition, 2015. Note: Large scale manufacturers operating at highest efficiency levels achieve lower costs (1/2 to 1/3 of cited cost).
Example Case: C1 for Insulin Production
C1 for mAb production shows promising initial results
Production of heavy & light chain successful MS/MS data reveals correct structure Binding to target confirmed via ELISA
Heavy chain
Light chain
Production of Fab successful The structure was confirmed by MS analysis Specificity of binding confirmed via ELISA
Lucentis
72h 96h Control
Case Study 1: Humira Case Study 2: Lucentis
C1 has produced biologically-active monoclonal antibodies Protease deficient strains with no mAb degradation successfully generated Codon use for fungal expression optimized Glycosylation controllable and glycoengineering is expected to begin in 2017
Successful Initial Engineering of C1 for mAb Production
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C1 production of mAbs could dramatically alter economics
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4218
124
60
110
360
0
50
100
150
200
250
300
350
400
C1 - 2000l tank C1 - 10,000l tank Standard Manufacturing
Annual OpEx
Initial CapEx Investment
Comparative Manufacturing Costs (Example: Humira for US market)
* OpEx cost include depreciation cost that assume depreciation of facility over 10 years, costs from active ingredient production only, no further processing1 requires two 2,000l tanks to satisfy annual production needs, 2 requires one 10,000l tank that will retain 10 months of production capacity, 3 requires three10,000l tanks
CHO - 10,000l tank 3
Cos
t in
Mill
ion
US
D
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Potential savings over current production methods warrant further engineering to realize
mAb production in C1
Biobetter mAbs Robust strain engineering and Glycoengineering
Novel mAbs
H1 2017 H2 2017 H1 2018 H2 2018
New Product Platform DevelopmentNew Product Development
mAb Biobetter Development
H1 2019
Estimated timeline for Further C1 Strain Engineering
*
C1 - 2,000l tank 1 C1 - 10,000l tank 2
C1 production is safe
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Non-Pathogenic
Not Genotoxic
Strain is non-toxic and non-infectious
No toxic byproducts are generated during production
In vivo trials demonstrated: No adverse effects No foreign DNA Safety confirmed
C1-cellulase accepted by FDA on September 29, 2009
GRAS notification letters are broadly recognized in the food and consumer products industries as the safety standard
For Enzyme Production: Generally Recognized as Safe (GRAS) Status
No Adverse Effects
Mice experiments showed: No adverse clinical
effects induced by C1 produced vaccines
Summary - Key Advantages of C1
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Dyadic is looking for partners in the biopharmaceutical space to exploit the potential of C1.
For further inquiry, please [email protected]
Further benefits: Unique properties that can be
engineered for the desired product profile
A toolbox for strain engineering to optimize production of different biologics (vaccines, simple proteins, antibodies)
Short production
cycles
2
High purity of produced
protein Robust and reliable
manufacturing
3
4
First product shown to be
safe in animal studies
5
Unprecedented protein yields
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