1
Update on
plant-made VLPs
Nathalie Landry
Vice President Product
Development
[email protected]
WHO meeting
May 6th 2014 © Medicago Inc.
All rights reserv ed
Medicago overview
2
Focus Vaccines & Protein-based pharmaceuticals
Manufacturing technology Transient expression in tobacco
Vaccine technology Virus-like particles
Employees 240
Headquarters, laboratories
& cGMP facilities
Quebec City, CANADA
Research Triangle Park, NC, USA
Genopole d’Evry, FRANCE
Corporate structure
Public company from 2006- Sept 2013
Private company since Sept 2013
Mitsubishi Tanabe Pharma Corporation
(majority shareholder)
© Medicago Inc.
All rights reserv ed
Transient expression in N. benthamiana
3
DNA sequence + plants Infiltration
Purification
Incubation
VLP Flu virus Extraction
© Medicago Inc.
All rights reserv ed
Influenza hemagglutinins accumulate at the plasma membrane
forcing curvature of the membrane and budding of virus-like particles
Influenza virus-like particle production in plants
From D’Aoust et al., Plant Biotechnology Journal, Volume 8: 607-619 (2010)
4
No need for NA:
No sialic acid in plants
2
© Medicago Inc.
All rights reserv ed
Rotavirus particle assembly
– Multi-step process initiated in the cytosol and completed in the ER
– Requires 4 structural antigens: VP2, VP6, VP7, VP4
Rotavirus-like particle production in plants
Plant-produced rotavirus-like particles
5 From Trask et al., Nature Reviews Microbiology, Volume 10: 165-177 (2012)
© Medicago Inc.
All rights reserv ed
Clinical trials – Pandemic vaccine
Phase I H5 VLP + Alhydrogel completed in 2009
– 48 healthy adults (18-60 years of age)
Phase II H5 VLP + Alhydrogel completed in 2010
– 255 healthy adults (18-60 years of age)
Phase I H5 VLP + GLA completed in 2013 (IDRI’s trial)
– 100 healthy adults (18-49 years of age)
– First trial evaluating an adjuvant with intradermal administration
– H5 VLP + Alhydrogel compared well with all other formulations
– GLA-AF increased cross-reactivity of antibodies
Phase II H5 VLP + GLA or Alhydrogel initiated in 2013
– 390 healthy adults (18-60 years of age)
– All subjects received 2 doses, no SAEs
– Analysis of immune response ongoing
Phase I H7 VLP + Alhydrogel initiated at beginning of 2014
– 100 healthy adults (18-60 years of age)
Clinical development overview and status update
6
Completed Ongoing
© Medicago Inc.
All rights reserv ed
Clinical trials – Seasonal vaccine
Phase I H1 VLP completed in 2011 (non-adjuvanted)
– 100 healthy adults (18-49 years of age)
– The 5 microgram dose met the CHMP criteria
– Good antibody response against human viruses
Phase I/II with seasonal quadrivalent launched in Oct. 2013
– 120 healthy adults (18-49 years of age)
Clinical development overview and status update
7
Completed Ongoing © Medicago Inc.
All rights reserv ed 8
From preclinical trials (one dose (10 µg), no
adjuvant):
– 100% protection in mice and ferrets against H5N1 Indonesia
– Independent NIAID heterologous challenge:
• 100% protection against H5N1 Vietnam
• 70% protection against H2N2 Japan
• Rec-HA failed to protect
Challenge against heterologous
H5N1 Vietnam
Challenge against heterolsubtypic
H2N2 Japan
Findings from preclinical studies Cross-protection
3
© Medicago Inc.
All rights reserv ed
**
* **
* #
GMT: 135 (16/16)
GMT: 27 (16/16)
GMT: 15 (12/16)
• 100% protection of mice immunized with one dose of 3 µg adj. H7 VLP
• 62.5% protection of mice immunized with one dose of 3 µg non-adj. H7 VLP
• 100% protection of mice immunized with 2 non-adj. doses of 3 µg (not shown)
• Phase I ongoing
*** significantly different from matched-placebo, p < 0.005
# significantly different from Placebo Alhydrogel, p < 0.05
Note: H7 VLP is not significantly different from adjuvanted
groups
Efficacy of H7 VLP after 1st dose in mice 100% protection against lethal challenge with H7N9
9 © Medicago Inc.
All rights reserv ed 10
H5 VLP
(A/Indonesia)
Medicago
H5N1 split
(A/Vietnam)
20 µg + 2.5µg GLA-AF (ID)
20 µg + 2.5µg GLA-AF (IM)
20 µg (ID) 20 µg + 0.5mg Alhydrogel(IM)
90 µg (IM)
Seroconversion rate
(4-fold increase) D42
Target of 40% 65.0% 80.0% 52.6% 83.3% 43.8%
Seroprotection rate
(% 1:32) D42
Target of 70%
70.0% 85.0% 52.6% 88.9% 50.0%
GMI D42/D0
Target of 2.5 10.3 8.7 4.9 11.4 4.5
H5 VLP with GLA-AF IM or ID or with alum IM meet the 3 CHMP criteria for
licensure of pandemic vaccines
H5 VLP clinical results Trial NCT01657929
© Medicago Inc.
All rights reserv ed
Phase I/II trial with seasonal quadrivalent VLP vaccine
11
TEST MATERIAL TREATMENT
GROUP
NO. OF
SUBJECTS DOSE LEVEL
ADMINISTERED ON
DAY
Seasonal quadrivalent VLP
Influenza vaccine
Treatment Grp 1
(low dose) 30
3 µg per
strain\ total of
12 µg HA
0
Seasonal quadrivalent VLP
Influenza vaccine
Treatment Grp 2
(medium dose) 30
9 µg per
strain\ total of
36 µg HA
0
Seasonal quadrivalent VLP
Influenza vaccine
Treatment Grp 3
(high dose) 30
15 µg per
strain\ total of
60 µg HA
0
100mM phosphate buffer + 150
mM NaCl + 0.01% Tween 80
Treatment Grp 4
(placebo) 30 None 0
© Medicago Inc.
All rights reserv ed
Safety profile Phase I/II with seasonal quadrivalent VLP vaccine (trial NCT01991587)
0%
10%
20%
30%
40%
50%
60%
70%
80%
90%
100%
Pla
ceb
o (
N=
30)
3 µ
g V
LP V
acci
ne
(n
=30)
9 µ
g V
LP V
acci
ne
(n
=30)
15
µg
VLP
Vac
cin
e (n
=30
)
Pla
ceb
o (
N=
30)
3 µ
g V
LP V
acci
ne
(n
=30)
9 µ
g V
LP V
acci
ne
(n
=30)
15
µg
VLP
Vac
cin
e (n
=30
)
Pla
ceb
o (
N=
30)
3 µ
g V
LP V
acci
ne
(n
=30)
9 µ
g V
LP V
acci
ne
(n
=30)
15
µg
VLP
Vac
cin
e (n
=30
)
PAIN ERYTHEMA SWELLING
Solicited local reactions, during the first 7 days after
immunization
Severe
Moderate
Mild
4
© Medicago Inc.
All rights reserv ed
Safety profile Phase I/II with seasonal quadrivalent VLP vaccine (trial NCT01991587)
0%
5%
10%
15%
20%
25%
30%
35%
40%
45%
50%
Pla
ceb
o (
N=
30)
3 µ
g V
LP V
acci
ne
(n
=30)
9 µ
g V
LP V
acci
ne
(n
=30)
15
µg
VLP
Vac
cin
e (n
=30
)
Pla
ceb
o (
N=
30)
3 µ
g V
LP V
acci
ne
(n
=30)
9 µ
g V
LP V
acci
ne
(n
=30)
15
µg
VLP
Vac
cin
e (n
=30
)
Pla
ceb
o (
N=
30)
3 µ
g V
LP V
acci
ne
(n
=30)
9 µ
g V
LP V
acci
ne
(n
=30)
15
µg
VLP
Vac
cin
e (n
=30
)
Pla
ceb
o (
N=
30)
3 µ
g V
LP V
acci
ne
(n
=30)
9 µ
g V
LP V
acci
ne
(n
=30)
15
µg
VLP
Vac
cin
e (n
=30
)
Pla
ceb
o (
N=
30)
3 µ
g V
LP V
acci
ne
(n
=30)
9 µ
g V
LP V
acci
ne
(n
=30)
15
µg
VLP
Vac
cin
e (n
=30
)
Pla
ceb
o (
N=
30)
3 µ
g V
LP V
acci
ne
(n
=30)
9 µ
g V
LP V
acci
ne
(n
=30)
15
µg
VLP
Vac
cin
e (n
=30
)
Pla
ceb
o (
N=
30)
3 µ
g V
LP V
acci
ne
(n
=30)
9 µ
g V
LP V
acci
ne
(n
=30)
15
µg
VLP
Vac
cin
e (n
=30
)
Pla
ceb
o (
N=
30)
3 µ
g V
LP V
acci
ne
(n
=30)
9 µ
g V
LP V
acci
ne
(n
=30)
15
µg
VLP
Vac
cin
e (n
=30
)
Pla
ceb
o (
N=
30)
3 µ
g V
LP V
acci
ne
(n
=30)
9 µ
g V
LP V
acci
ne
(n
=30)
15
µg
VLP
Vac
cin
e (n
=30
)
Pla
ceb
o (
N=
30)
3 µ
g V
LP V
acci
ne
(n
=30)
9 µ
g V
LP V
acci
ne
(n
=30)
15
µg
VLP
Vac
cin
e (n
=30
)
FATIGUE HEADACHE MUSCLESACHES
JOINT ACHES CHILLS FEVER GENERALDISCOMFORT
SWELLING INTHE AXILLA
SWELLING INTHE GROIN
SWELLING INTHE NECK
Solicited systemic reactions, during the first 7 days after
immunization
Severe
Moderate
Mild
OT over 38C
© Medicago Inc.
All rights reserv ed
Phase I/II with Quadrivalent VLP vaccine Hemagglutination-Inhibition Antibody Titers 21 days post-dose (trial NCT01991587)
14
Strain Criteria Groups
3 µg/strain 9 µg/strain 15 µg/strain Placebo
H1N1
≥ 4-fold (%) 41.4 50.0 40.7 0
≥ 1:40 (%) 79.3 73.3 81.5 33.3
GMI ≥ 2.5 (%) 3.6 4.5 3.7 1.1
H3N2
≥ 4-fold (%) 48.3 60.0 44.4 0
≥ 1:40 (%) 79.3 90.0 81.5 23.3
GMI ≥ 2.5 (%) 4.7 7.6 5.0 1.0
B/Brisbane
≥ 4-fold (%) 13.8 43.3 48.1 3.3
≥ 1:40 (%) 65.5 70.0 85.6 30.0
GMI ≥ 2.5 (%) 2.0 3.9 3.8 1.1
B/Wisconsin
≥ 4-fold (%) 24.1 53.3 51.9 10.0
≥ 1:40 (%) 72.4 73.3 96.3 43.3
GMI ≥ 2.5 (%) 2.3 6.3 4.9 1.3
Meets CHMP criteria
© Medicago Inc.
All rights reserv ed
Phase I/II with Quadrivalent VLP vaccine Micro-Neutralisation antibody titers 21 days post-dose (trial NCT01991587)
15
Strain Criterium Groups
3 µg/strain 9 µg/strain 15 µg/strain Placebo
H1N1
GMT 93.4 103.1 153.9 16.2
≥ 4-fold (%) 44.8 46.7 55.6 0
H3N2
GMT 436.6 544.4 570.2 62.0
≥ 4-fold (%) 48.3 70.0 63.0 0
B/Brisbane
GMT 24.5 31.0 57.3 13.0
≥ 4-fold (%) 20.7 26.7 40.7 0
B/Wisconsin GMT 69.3 71.3 107.5 23.0
≥ 4-fold (%) 34.5 53.3 44.4 0
B/Mass (Yamagata strain for 2014-2015
season)
GMT 27.9 30.6 62.6 12.7
≥ 4-fold (%) 13.8 26.7 33.3 0
© Medicago Inc.
All rights reserv ed
A single dose of H1 VLP vaccine induced polyfunctional T cell
response PBMCS collected 6-month post-vaccination, trial NCT01302990
Peptide pool stimulation H1 H1 VLP stimulation
Placebo H1 VLP Fluzone Pie CategoryTest Results
Placebo H1 VLP Fluzone
Placebo
H1 VLP 0.0042
Fluzone 0.8281 0.0276
Placebo H1 VLP Fluzone Placebo H1 VLP Fluzone
Re
sp
on
siv
e C
D4
+ t
o H
1 p
ep
t.
po
ol p
er
10
6 C
D4
+ T
ce
lls
Re
sp
on
siv
e C
D4
+ t
o H
1 p
ep
t.
po
ol p
er
10
6 C
D4
+ T
ce
lls
IFN-g
IL-2
TNF-a
+
+
+
-
+
+
+
-
+
+
+ -
-
+ -
+
- -
Pie Chart Arc Legend
IFN-g +
IL-2 +
TNF-a +
Arc legend
5
© Medicago Inc.
All rights reserv ed
H1 VLP induces cross-reactive T cells to H5N1 PBMCs collected 6 months after vaccination, trial NCT01302990
• More CD4 & CD8 T cells cross-reactive for H5N1 compared to
egg-based and placebo
• Cross-protective effects similar to those reported for H5N1
vaccines administered with oil-in-water adjuvants 17
Cross-reactive responses of CD4+ (left panel) and CD8+ (right panel) T cell responses from the H1 VLP-
vaccinated patients ex vivo stimulated with H5 VLP. The symbol * indicates statistically significant
differences between two-groups analyzed by the Mann-Whitney test (P≤0.05).
© Medicago Inc.
All rights reserv ed Cytokines expressing profile in CD4+ cells after ex vivo stimulation with H5 peptides pool 18
IL-2
IFN-g
TNF-a
+
+
+
+
+
-
+
-
+
+
-
-
-
+
+
-
+
-
-
-
+
Re
sp
on
siv
e C
D4+
ce
lls
to H
5 p
ep
tid
es p
er
10
6 C
D4+
ce
lls
10 µg H5 VLP + Alhy
20 µg H5 VLP + Alhy
15 µg H5 VLP + Alhy
Placebo
Polyfunctional CD4+ T Cell response to H5 Homotypic response after vaccination with H5 VLP (21 days post-boost, trial
NCT01991561)
* *
*
*
Statistically significant
compared
to placebo
*
© Medicago Inc.
All rights reserv ed
Cross-reactive CD4+ T Cell response to HA of H2N2 Heterotypic (group 1) response after vaccination with H5 VLP (trial
NCT01991561)
IL-2
IFN-g
TNF-a
+
+
+
+
+
-
+
-
+
+
-
-
-
+
+
-
+
-
-
-
+
Placebo
Re
sp
on
siv
e C
D4+
ce
lls
to H
2 V
LP
pe
r 10
6 C
D4+
ce
lls
10 µg H5 VLP + Alhy
20 µg H5 VLP + Alhy
15 µg H5 VLP + Alhy
19
One non-adj
H5 VLP dose:
70% protection
against H2N2
in mice
Cytokines expressing profile in CD4+ cells after ex vivo stimulation with H2 VLP
* * *
Statistically significant
compared
to placebo
*
© Medicago Inc.
All rights reserv ed
Cross-reactive CD4+ T Cell response to HA of H7N9 Heterotypic (group 2) response after vaccination with H5 VLP (trial
NCT01991561)
Re
sp
on
siv
e C
D4+
ce
lls
to H
7 V
LP
pe
r 10
6 C
D4+
ce
lls
IL-2
IFN-g
TNF-a
+
+
+
+
+
-
+
-
+
+
-
-
-
+
+
-
+
-
-
-
+
10 µg H5 VLP + Alhy
20 µg H5 VLP + Alhy
15 µg H5 VLP + Alhy
Placebo
20 Cytokines expressing profile in CD4+ cells after ex vivo stimulation with H7 VLP
*
*
Statistically significant
compared
to placebo
*
6
© Medicago Inc.
All rights reserv ed
H5 VLP pandemic vaccine When formulated with alum, induce HI antibody titers that meet surrogate
correlates of protection based on HI antibody titers
Induce polyfunctional T cell response
H7 VLP pandemic vaccine Induced a comparable or superior antibody response to H5 VLP in 2 animal
models
Currently evaluated in humans formulated with alum
Seasonal Quadrivalent vaccine – Good safety profile
– Dosages of 9 and 15 µg per strain meets the CHMP criteria for licensure of seasonal vaccines
– Induced a good MN antibody response cross-reactive towards other strain (B Yamagata)
Conclusion
21 © Medicago Inc.
All rights reserv ed
Animal challenge studies and CMI results during clinical trials with plant-based VLP vaccines show potential for cross-protection
– H5 vs. H1, H2 & H7 demonstrated to date
CMI important for seasonal flu vaccine in terms of cross-protection – 2009 H1N1 pandemic: elderly protected by memory CD4 response from previous
H1N1 infections (Hsu et al. 2012 Int J Infect Dis)
– Shown to protect humans in the absence of antibodies (Wilkinson 2012)
– Characterization ongoing in current Phase I/II with seasonal VLP vaccine
Plant-based VLPs – Numerous manufacturing advantages
– Good safety and immunogenicity results in ~1,000 subjects
– Good antibody response
– Cross-protective effects possibly related to strong T cell response
– Good compromise to natural infection
Conclusion
22
© Medicago Inc.
All rights reserv ed
To evaluate the alum-adjuvanted pandemic vaccine in a pivotal phase 3 trial To increase the safety database
Will include a lot-to-lot consistency trial
To continue the clinical evaluation of the seasonal quadrivalent VLP
vaccine
Larger group size to evaluate CBER criteria for licensure
In healthy adults
In the elderly population
Future plans
23 © Medicago Inc.
All rights reserv ed
Merci!
Thank you!
Arigatō!
