Novozymes Veltis® is a clinically-proven half-life extension technology based on engineered albumins that allows drug developers to optimize dose size and frequency to achieve improved patient compliance.

1. VELTIS® Engineered albumins for optimized drug dosing
Dr Darrell Sleep, PEGS - Peptide Therapeutics, 8th May 2014
Designed by Nature. Perfected by Novozymes.

2. VELTIS® - INNOVATIVE TECHNOLOGY ENABLING
YOU TO RETHINK YOUR THERAPEUTIC WINDOW
 Veltis® is an albumin based drug
delivery technology that improves
drug efficacy and patient
compliance based upon the natural
transportation properties of albumin
 Veltis® delivers control over dose
frequency, dose quantity and drug
tolerability using engineered human
albumin
 Approved in GSK’s Eperzan*/
Tanzeum**, Veltis® delivers
outstanding performance from the
albumin experts
 Science
 Knowledge
 Expertise
 IP
*EU Approval in March 2014, **FDA Approval in April 2014
All you need to do is define
your dosing regime

3. VELTIS® TECHNOLOGY PIPELINE
Drug
Pre-clinical
Phase I Phase II
Phase
III
BLA /
MAA
Submitted
Approved
Diabetes
Neutropenia
Haemophilia
WT albumin
Engineered
Albumins
“… we believe it offers unique product and patient benefits to
enhance our current protein therapeutic pipeline… pre-clinical
studies are encouraging.”
Eperzan
Tanzeum
Balugrastim
Factor-IX
Unnamed
T-regs

4. Albumin has a naturally long plasma half-life:
 Human albumin – T1/2~19 days
 Size - retained by kidney/glomerulus
 FcRn (neonatal Fc receptor) recycling: pH-dependent recycling “rescues”
albumin from degradation and prolongs half-life
ALBUMIN IS RESCUED FROM DEGRADATION BY
THE FcRn RECEPTOR

5. Albumin has a naturally long plasma half-life:
 Human albumin – T1/2~19 days
 Size - retained by kidney/glomerulus
 FcRn (neonatal Fc receptor) recycling: pH-dependent recycling “rescues”
albumin from degradation and prolongs half-life
60%
Albumin + FcRn
engagement
Albumin - FcRn
engagement
ALBUMIN IS RESCUED FROM DEGRADATION BY
THE FcRn RECEPTOR

6. Time
Serum Concentration
 The pharmacokinetics of albumin
fusions and conjugates are typically
lower than that of albumin alone
 Final drug product can be cleared
through either the albumin or the
drug component
 Improved control over the final half-life
of albumin fusion or conjugate to
achieve monthly, two-weekly, or
weekly peptide or protein drug
dosing
 Define therapeutic dosing window by
balancing dose size and frequency to
achieve optimum efficacy and
tolerability
Albumin
Albumin
Fusion
Free
Drug
WHY ENGINEER HUMAN ALBUMIN?
Illustrative PK curves

7. DI
DIII
FcRn
C-TERMINAL REGION OF DIII IS IMPORTANT
FOR FcRn BINDING
H464 D494
H535
H510
C-terminal
a-helix
C-terminal truncatio1n0 8o-f1 a1l2b umin
led to a nine-fold reduction in
FcRn binding affinity

8. DI
DIII
FcRn
CONSERVED HISTIDINES IN DIII ARE
IMPORTANT FOR FcRn BINDING
H464 D494
H535
H510
C-terminal
a-helix
H440 does not alter 1F0c8R-n11 b2i nding
(least conserved)

9. DI
DIII
FcRn
COMPUTATIONAL MODELLING IDENTIFIES DI
AS IMPORTANT FOR FcRn BINDING
H464 D494
H535
H510
C-terminal
a-helix
78-88
108-112
Loop predictions from docking
model1 interact in the FcRn co-crystal
structure (Schmidt et al.
(2013))
1. Andersen, J. T. et al. Structure-based mutagenesis reveals the albumin-binding site of the neonatal Fc
receptor. Nature Communications 3, 610 (2012)

10. RECEPTOR AFFINITY CAN BE TUNED UP OR
DOWN …WITH A SINGLE POINT AMINO ACID
MUTATION

11. ENGINEERING THE ALBUMIN-FcRn INTERACTION
FOR ENHANCED PHARMACOKINETICS
 Albumins have been engineered
for increased and decreased
binding affinity to the human
FcRn receptor
 Variants selected for
enhanced binding at
endosomal pH and no
significant binding at
neutral pH
 FcRn binding affinity can be both
increased and decreased in a pH
dependent manner
 Variants contain between 1-5
amino acid substitutions

12.  In silico studies
 EpiMatrix Protein score
 -29.9 to -32.0
 In vitro Class II HLA binding
and ex vivo T-cell assays
 No significant HLA binding
2nd
generation
albumin
variants
ENGINEERED ALBUMINS - NO SIGNIFICANT
IMMUNOGENICITY PREDICTED
- 80 -
- 70 -
- 60 -
- 50 -
- 40 -
- 30 -
- 20 -
- 10 -
- 00 -
- -1 0 -
- -2 0 -
- -3 0 -
- -4 0 -
- -5 0 -
- -6 0 -
- -7 0 -
- -8 0 -
Thrombopoietin
Erythropoietin
IgA
Fibrinogen-Gamma
Albumin
IgG FC Region
GMCSF
Follitropin - Beta
Fibrinogen-Alpha
Beta-2-Microglobulin
Interferon-Beta
GHRH
Tetanus Toxin
Influenza-HA
EpiMatrix Scores
“Considering our global and
regional analysis as a whole,
we find the risk that these
proteins will create or
contribute to anti-therapeutic
immune response to be
minimal”

13. WT Mice and Tg Mice
Macaque
Rodents
Cross species
binding difference
Single
transgenic
mice
Human
receptor/Mouse
albumin
Primate
Closest to humans
In vitro receptor
binding
TRANSLATING FcRn BINDING TO IN VIVO
PHARMACOKINETICS
How does Veltis® behave in
common animal species?

14. Animal FcRn affinities
 Implications for PK and
transgenic animals
Wild type human albumin
(fusions) will be out competed by
MSA in mouse model
RODENT PK STUDIES
– NOT AS SIMPLE AS YOU MIGHT THINK
KD (μM) HSA MSA
Human FcRn 4.5 0.8
Mouse FcRn 86 9.3
Albumin Model T1/2(h)
Human Rat 15
Rat Rat 49
Human albumin has a
short half-life in rats
Explained by cross-species FcRn
binding properties
 Human albumin binds very
poorly to FcRn from rodents
Andersen et al. (2010) J. Biol. Chem. 285(7):4826-36

15. FcRn BINDING TRANSLATES TO PK MODIFICATION
IN WILD TYPE MICE
V0098 - Improved FcRn
binding variant
 Half-life extension (31h)
 Increase in AUC
 Reduced clearance
 Increased FcRn rescue
V0088 -Reduced FcRn
binding variant
 Shorter half-life (19h)
 Decrease in AUC
 Increased clearance
 Reduced FcRn rescue
Animal model: WT NMRI Mice; Single bolus intravenous administration; 10mg/kg
V0098 - T½ 31h
WT - T½ 21h
V0088 – T½ 19h

16. Animal model: human FcRn transgenic mouse (Tg32); Single bolus intravenous administration; 10mg/kg
V0098 - Improved FcRn
binding variant
 Half-life extension (95h)
 Increase in AUC
 Reduced clearance
 Increased FcRn rescue
V0088 - Reduced FcRn
binding variant
 Shorter half-life (31h)
 Decrease in AUC
 Increased clearance
 Reduced FcRn rescue
V0098 - T½ 95h
WT - T½ 67h
V0088 – T½31h
FcRn BINDING TRANSLATES TO PK MODIFICATION
IN HUMAN FcRn TRANSGENIC MICE

17. 2 subjects/compound, 1mg/kg dose, Sampling: pre-dose-50 days (1200 hr)
ENGINEERED ALBUMINS ACHIEVE MORE THAN A
DOUBLING OF HALF-LIFE IN A PRIMATE MODEL
 Veltis albumins V0098,
V0354 and V0311 show a
1.6, 2.1 and 2.4 longer half-life,
respectively, compared
to WT human albumin
 The prolongation in half-life is
reflected in a reduced
clearance, increased mean
residence time (MRT) and
increased AUC
 Doubling of half-life in
primates opens the door to
monthly dosing of
therapeutic peptides and
proteins in humans

18.  The pharmacokinetics of the
variant albumins were
evaluated in a number of in
vivo models
 WT Mouse
 Transgenic mouse*
 Non human primate
 Albumin variants show similar
trends across species with
increased and decreased FcRn
binding relating directly to
increased and decreased half-life
 Small differences observed
between variants in the WT
mouse model are due to
 Low affinity of human albumin
to the mouse FcRn receptor
 Competition for FcRn binding
with the endogenous mouse
albumin
SUMMARY OF VELTIS® PK DATA FROM A RANGE
OF IN VIVO MODELS
NT – Not tested
*Transgenic mouse – human FcRn in mouse albumin background
NT NT NT

19. RECEPTOR AFFINITY MAINTAINED WHEN DRUGS
ARE FUSED OR CONJUGATED TO VELTIS®
ALBUMINS
Test Construct - fusion
hFcRn
KD
(mM)
WT Albumin 3.1
WT-FLAG 2.9
scFv-WT-scFv-FLAG 2.0
V0098 0.1
V0098-FLAG 0.2
V0098-IL1ra 0.2
scFv-V0098-FLAG 0.2
scFv-V0098-scFv-FLAG 0.1
V0098-scFv-FLAG 0.1
scFv-V0098 0.1
Test Construct - conjugate
hFcRn
KD
(mM)
WT Albumin 4
WT Albumin+Exendin-4 5
V0098 0.4
V0098+Exendin-4 0.5
V0354 0.2
V0354+Exendin-4 0.2

20. PHARMACOKINETIC IV PROFILES OF ALBUMIN
EXENATIDE CONJUGATES IN WT MICE
Variant AUC (h.ug/mL) Cl (ml/h/kg) Vz (mL/kg) t½ (h) t½ fold increase
WT-HSA 714.88 6.99 97.4 10.9 -
V0098 851.9 5.87 109.3 12.9 1.2
V0354 958.38 5.22 114.9 15.3 1.4

21. PHARMACODYNAMICS OF ALBUMIN EXENATIDE
CONJUGATES IN NONFASTED DIABETIC MICE

22. COMPLETE SUPPORT PACKAGE FROM THE
ALBUMIN EXPERTS
Rapid Proof of Concept
 Fusion and conjugate design
 Open research licences
 Veltis® albumin samples
Veltis® Tool Box
 FcRn and cell assays
 Yeast expression
 Transgenic animal
 Conjugation technology
Clinical Development
 Phase I-II supply through Novozymes
or partners
 Tech transfer to CMO
 cGMP supply of Veltis® albumins
Pre-clinical Development
 Fermentation optimisation
 Process development
 Provision of materials for
toxicology and in vivo studies
from Novozymes or partner

23. We provide the definitive technology
You define your therapeutic window
VELTIS®
Designed by Nature. Perfected by Novozymes.
Proven clinical performance
Patient-friendly dosing and low risk of adverse
events
Engineered albumins for optimized drug
dosing with significant half-life extension over
native albumin
Long patent life
Flexible and scalable manufacturing by fusion
or conjugation
Provided by technology developer with strong
heritage and track record in albumin supply

24. THANK YOU
CONTACT:
DSEE@NOVOZYMES.COM
Booth No: 200
Poster No: 318