Delivery of ALX-0171 by inhalation greatly reduces disease burden in a neonatal

Delivery of ALX-0171 by
inhalation greatly reduces
disease burden in a neonatal
lamb RSV infection model
9th RSV Symposium
Stellenbosch –South Africa
13th November 2014
Laurent Detalle
Nanobodies® Inspired by nature
Disclosure
Laurent Detalle
• employee at Ablynx
Study performed at Iowa State University
• paid for by Ablynx
• Ablynx as study monitor
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2
Ablynx – Facts and figures
Corporate
• Drug discovery and development company in Ghent, Belgium
• >300 employees
Technology
• Pioneer in next generation biologics – Nanobodies®
• >500 granted and pending patents
Products
• >30 programmes – six in clinical development
• Three clinical proof-of-concepts (POC)
• >900 healthy volunteers and patients treated with Nanobodies
Partners
• AbbVie, Boehringer Ingelheim, Eddingpharm, Merck & Co,
Merck Serono and Novartis
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Nanobodies – derived from heavy-chain only antibodies
Camelid heavy-chain only antibodies are stable and fully functional
Nanobodies represent the next generation of antibody-derived biologics
VH
VHH
CH1
VHH
VL
CL
CH2
CH3
Conventional
antibodies
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Ablynx’s Nanobody
CH2
CH3
Heavy chain only
antibodies
• small
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• sequence homology comparable
to humanised/human mAbs
• easily linked together
• nano- to picomolar affinities
• intractable targets
• multiple administration routes
• manufacturing in microbial cells
4
ALX-0171 – designed to treat infant RSV infection
First-in-class treatment of RSV infection
Being developed to treat hospitalised and out-patient infants
anti-RSV
Nanobody
anti-RSV
Nanobody
Trivalent Nanobody: demonstrated in vitro efficacy against A
and B serotypes
ALX-0171
42kD
anti-RSV
Nanobody
Pulmonary delivery: fast onset of action and high concentration at
infection site
In vivo efficacy demonstrated in cotton rat model and neonatal lambs
Three phase I studies in man with inhaled ALX-0171 successfully
completed
First-in-infant phase IIa study expected to start in Nov/Dec 2014
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5
ALX-0171 – translational strategy
Bridging the translational gap between adults and infants
Requirement for a fit-for-purpose model
ALX-0171
Efficacy/safety
•
ALX-0171
delivery
Preclinical
In vitro studies
Preclinical
In vivo studies
ALX-0171
characterisation
Efficacy in cotton rats
Safety and PK
in adult/juvenile rats
Stability/
nebulisation
studies
Intranasal/whole
body delivery
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Clinical
In vivo studies
Safety and PK in
Human adults
Oral inhalation
Breath
actuation
Preclinical
In vivo studies
Efficacy,
Safety and PK in
Neonatal setting with
relevant administration
route
Nasal
inhalation with
facemask
6
Neonatal lambs as model system for safety and efficacy
testing of inhaled ALX-0171 upon RSV infection
Safety
Age/lung
development
• 2-5 days old
• Colostrumdeprived
• Alveolarisation
starts preterm1
• Immature
immune system
Lung deposition/
Pharmacokinetics
Size
• Allows relevant
administration
route (facemask)
• Enables multiple
assessments
• Similar breathing
patterns
Respiratory
tract anatomy
• Size of nasal
cavity and
airways2
• Organisation of
local lymphoid
tissue2
• Airway branching
pattern2
• Distributon of
epithelial cells,
mast cells and
airway smooth
muscle3
• ...
Efficacy
RSV disease
• Lower
respiratory tract
infection
• Develop mild
clinical
symptoms4
• Histologic
lesions4
• Enhanced
disease postvaccination5
Similar characteristics to human infants
1ATS
4Derscheid,
2Scheerlinck,
5Derscheid,
American journal of respiratory and critical care medicine, 2004. 170(3): p. 319-43
J.P., et al., Trends in biotechnology, 2008. 26(5): p. 259-66.
3Meeusen, E.L., et al., Drug Discovery Today, 2009. 6(4): p. 101-106
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R.J. and M.R. Ackermann, Viruses, 2012. 4(10): p. 2359-78.
R.J., et al. PloS one, 2013. 8(12): p. e81472.
7
Lamb study design
Objectives
• Assess efficacy/safety in neonatal setting
Nebulisation of
RSV M37 strain or
culture medium
Necropsy
Onset of
RSV infection
- therapeutic setting starting at viral peak
• Pharmacokinetics in target tissue
- epithelial lining fluid (ELF)
Day -1
0
1
2
3
4
5
6
7
8
5-6 animals/group
Note: Pilot study was performed with similar schedule
Treatment
ALX-0171 or formulation buffer
Study design
Group
RSV
status
Treatment
Nebuliser
filling dose
Time taken
for dose to
be delivered
Endpoints
1
Mockinfected
Vehicle
-
-
2
Mockinfected
ALX-0171
11 mg
(Low dose)
~2 minutes
•
•
•
•
•
Viral titers (qPCR, FFU)
Viral antigen expression in lungs (IHC)
Gross lung viral lesions
Histopathology
General health status
3
RSV
Vehicle
-
-
4
RSV
ALX-0171
11 mg
(Low dose)
~2 minutes
•
ELF Ctrough ALX-0171 levels
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8
Nebulisation and PK
Nebuliser
• Mesh nebuliser
• MMAD: 3.27 ± 0.13 μm
• 2L/min airflow
A L X - 0 1 7 1 c o n c e n t r a t io n s i n
A L X - 0 1 7 1 c o n c e n t r a t io n s
lu n g s
1000
100
L o w d o s e A L X -0 1 7 1
H ig h d o s e A L X - 0 1 7 1
100
IC 9 0
LLO Q
[ A L X - 0 1 7 1 ] E L F ( µ g /m L )
10
1
0 .1
IC 9 0
o
L
d
A
L
o
w
V
S
R
s
X
e
V
V
S
R
e
h
-0
1
ic
7
le
1
7
e
1
s
-0
o
d
X
L
o
w
A
L
N o m in a l t im e ( h o u r s p o s t f ir s t d o s e )
80
le
60
ic
40
h
20
e
0
1
0 .0 1
10
V
[ A L X -0 1 7 1 ] p la s m a ( n g /m L )
in p la s m a f r o m p ilo t s t u d y
ALX-0171 concentrations in ELF were far above the IC90 following 3 daily
inhalations and 2-3 log higher than systemic concentrations
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Improved health status in RSV infected lambs
General health status of lambs was scored on a daily basis
G e n e r a l i ll n e s s s c o r e
Score
General illness score
0
No clinical signs
1
Reluctant to move
2
Reluctant to move, head down, depressed, not
interested in eating
3
Down, unwilling to get up or difficulty standing, not
eating
4
Down and should be euthanised, probably cannot eat
% la m b s w it h s c o r e  1
100
-1
M o c k V e h ic le
M o c k A L X -0 1 7 1
80
R S V V e h ic le
R S V A L X -0 1 7 1
60
40
20
0
1
2
3
4
5
6
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6
5
4
3
2
1
y
a
D
D
a
y
8
6
4
a
D
D
a
y
y
3
1
0
y
Poster number 120
n e o n a ta l la m b s
a
Treatment with ALX-0171 treatment improved the health
status
V ir a l k in e t ic s i n
D
Decline in general health status was observed in all lambs
starting on day 3
V ir a l t it e r s in B A L F ( L o g 1 0 F F U /m L ) (  s e )
D a y s p o s t in f e c tio n
10
Antiviral effect of ALX-0171
ALX-0171 decreases viral load in the lungs of infected lambs
Viral Titers
V ir a l t it e r s
 4 .1 L o g
6
4
2
DL
1
1
7
le
-0
ic
X
h
L
e
A
Viral RNA
V ir a l R N A in B A L F
S
R
R
S
V
V
A
V
L
e
X
h
-0
ic
1
7
le
1
1
7
1
-0
X
L
A
1
-0
X
L
A
V
S
R
0
le
1
7
le
h
e
V
R
S
V
A
L
e
X
h
-0
ic
1
ic
7
le
1
0
2
ic
2
h
4
4
e
6
 0 .4 7 L o g
6
V
8
L o g 1 0 M 3 7 R N A c o p ie s /m g lu n g (  s e )
V ir a l R N A in lu n g t is s u e
 0 .5 5 L o g
V
L o g 1 0 M 3 7 R N A c o p ie s /m L (B A L ) (  s e )
R
R
S
S
V
V
A
V
L
e
X
h
-0
ic
1
7
le
1
0
V
L o g 1 0 F F U /m L B A L ( R t C d lo b e ) (  s e )
• 4 log10 on cultivatable virus
• 0.5 log10 on viral RNA copies (>1 Log10 in pilot study)
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Antiviral effect of ALX-0171
ALX-0171 decreases viral load in the lungs of infected lambs
RSV Vehicle
10
5
1
7
1
X
L
A
V
S
R
R
S
V
A
L
V
X
e
h
-0
-0
1
ic
7
le
1
0
X
h
-0
ic
1
7
le
1
1
7
1
-0
X
1
-0
X
L
15
0
le
1
7
le
h
e
V
L
e
A
S
R
R
S
V
V
A
V
L
e
V
A
R
S
V
S
R
20
Daily inhalation of ALX-0171 markedly reduced viral titres and lung
lesions in RSV-infected lambs
V
A
L
e
X
h
-0
ic
1
ic
7
le
1
0
2
ic
2
25
Decrease in virus load is reflected
by a strong decrease in viral antigen
expression
4
h
4
L o g 1 0 M 3 7 R N A c o p ie s /m g lu n g (  s e )
6
 0 .4 7 L o g
6
80
le
V
S
S
R
R
V ir a l R N A in lu n g t is s u e
 0 .5 5 L o g
V
L o g 1 0 M 3 7 R N A c o p ie s /m L (B A L ) (  s e )
V ir a l R N A in B A L F
A lv e o li
100
ic
-0
X
L
A
Viral RNA
B r o n c h i/b r o n c h io le s
120
h
1
1
7
le
ic
h
e
V
A
V
L
e
X
h
-0
ic
1
7
le
1
0
140
e
DL
IH C s c o r e s
V ir a l a n t i g e n e x p r e s s io n
V
2
M e a n n u m b e r o f a f fe c te d
4
b r o n c h i/b r o n c h io le s o r a lv e o li p e r fie ld (  s e )
 4 .1 L o g
8
RSV ALX-0171
Viral Titers
V ir a l t it e r s
6
V
L o g 1 0 F F U /m L B A L ( R t C d lo b e ) (  s e )
• 4 log10 on cultivatable virus
• 0.5 log10 on viral RNA copies (>1 Log10 in pilot study)
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Effect of ALX-0171 on viral lung lesions
Plum red RSV lesions seen in lungs of RSV infected lambs on day 6
post-infection
• present on all lung lobes assessed
R ig h t C ra n ia l
R ig h t M id d le
50
R ig h t C a u d a l
A cc e s so ry
40
L e ft C r a n ia l
30
L e ft M id d le
L e ft C a u d a l
20
10
-0
X
L
A
S
R
R
S
V
V
A
V
L
e
X
h
-0
ic
1
7
le
7
1
ic
h
e
V
1
1
0
le
M e a n % in v o lv m e n t /lo b e (  s e )
G r o s s v ir a l le s io n s
Daily inhalation of ALX-0171 markedly reduced gross lung viral lesions
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Histological scoring of lamb lung
Alveolar consolidation was scored
Observed lesions
inflammatory infiltrate in bronchiolar lumen, alveolar spaces, and alveolar septa
Bronchioles and alveolar spaces variably filled with degenerate neutrophils and
sloughed epithelial cells with occasional large epithelial syncytial cells.
0% consolidation = 0
1%-9% consolidation = 1
10%-39% consolidation = 2
40%-69% consolidation = 3
70%-100% consolidation = 4
2
1
1
7
L
A
V
S
R
R
S
V
A
L
V
X
e
h
-0
1
ic
7
-0
X
h
e
V
Degenerate/Necrotic epithelial cells
le
1
0
ic
Syncytial cell
3
1
Accumulation of degenerate neutrophils
4
le
alveolar septa mildly to moderately thickened by hyperplasia of type II cells
H is to lo g ic a l c o n s o lid a t io n s c o r e
alveolar spaces variably filled with degenerate neutrophils and sloughed epithelial
cells
Daily inhalation of ALX-0171 markedly reduced microscopic lung viral
lesions
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ALX-0171 – Conclusions
Neonatal lambs proves to be a relevant model for treatment of RSV infection
Administrations of nebulized ALX-0171 resulted in effective target lung
concentrations and low systemic exposure
ALX-0171 administration was well tolerated with no adverse events noted in a
neonatal setting
ALX-0171 decreased RSV disease burden in neonatal lambs when treatment
started at peak of viral load and when symptoms were apparent
• Beneficial influence shown on:
- general health status
- viral titers and lung viral antigen expression
- gross and microscopic lung viral lesions
First-in-infant study expected to start in Nov/Dec 2014 using a customdeveloped infant inhalation device based on a vibrating mesh
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15
Acknowledgements
Ablynx, Gent, Belgium
•
•
•
•
•
•
•
•
•
•
•
•
•
•
•
Thomas Stöhr
Kjell Mortier
Linde Duprez
Lieselot Bontinck
Massimiliano Germani
Judith Baumeister
Sandy Jacobs
Donata Thuy
Toon Wauman
Gregory Daelman
Koen Allosery
Erik Depla
Catelijne Stortelers
Stephanie Staelens
Francis Descamps
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Iowa State University
•
•
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Mark R Ackermann
Jack M Gallup
Albert Van Geelen
Alejandro Larios-Mora
Shannon Jones Hostetter
IWT, Belgium
• Grant 130562
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