novozymes veltis® – engineerd albumins for optimized drug dosing

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VELTIS® Engineered albumins for optimized drug dosing

Dr Darrell Sleep, PEGS - Peptide Therapeutics, 8th May 2014

Designed by Nature. Perfected by Novozymes.

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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

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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

http://www.janssen.com/index.html

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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

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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

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Time

Seru

m C

oncentr

ation

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

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DI

DIII

FcRn

C-TERMINAL REGION OF DIII IS IMPORTANT FOR FcRn BINDING

H464 D494

H535

H510

C-terminal a-helix

108-112 C-terminal truncation of albumin led to a nine-fold reduction in

FcRn binding affinity

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DI

DIII

FcRn

CONSERVED HISTIDINES IN DIII ARE IMPORTANT FOR FcRn BINDING

H464 D494

H535

H510

C-terminal a-helix

108-112 H440 does not alter FcRn binding (least conserved)

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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)

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

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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

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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 -

- - 10 -

- - 20 -

- - 30 -

- - 40 -

- - 50 -

- - 60 -

- - 70 -

- - 80 -

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”

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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?

http://www.google.co.uk/url?sa=i&rct=j&q=&esrc=s&frm=1&source=images&cd=&cad=rja&docid=MU8csyLRTnlAVM&tbnid=9QZ-cK84VWpwJM:&ved=0CAUQjRw&url=http://www.fefchemicals.com/biopharm/scientific-information/articles/&ei=76akUdTAAYKe0QXBo4GgAg&bvm=bv.47008514,d.bGE&psig=AFQjCNGlRr2HCHZK1IRs5ngq6BEEbIUkog&ust=1369831530395613

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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

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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

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guide 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

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guide 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

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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

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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

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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

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PHARMACODYNAMICS OF ALBUMIN EXENATIDE CONJUGATES IN NONFASTED DIABETIC MICE

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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

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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

THANK YOU CONTACT: [email protected]

Booth No: 200 Poster No: 318

novozymes veltis® – engineerd albumins for optimized drug dosing

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