Transcript Development of Advanced Adjuvants and Immune Modulators

Development of Advanced Adjuvants
and Immune Modulators
100
Therapeutic IDR-1018 +
Anti-malarial
10 6 PbA
90
70
% Survival
40,000
p = 0.034
80
30,000
d0
4 days (Par 3-6%)
60
Anti-malarial 
Control Peptide
Pyrimethamine +
Chloroquine in
drinking water
50
40
Immunize
20,000
30
10,000
20
1018 or
IDR-1
d 4, 5, 6
10
No Treatment
0
0
0
1
2
3
4
5
6
7
8
9
10 11 12 13
Day 0
Days post-infection
R.E.W. (Bob) Hancock,
Centre for Microbial Diseases and
Immunity Research,
University of British Columbia
Day 22
Cationic Peptides
Birds do it, bees do it, even
educated fleas do it.... Cole Porter
 Important host defense mechanism in all
complex species of life.
 >1000 peptides known. Diverse amino acid
sequences and structures.
 12 to 40 (or more) amino acids. Net charge
+2 to +9 (Lys; Arg). Amphipathic.
 Role in Innate Immunity involves
antimicrobial, immunomodulatory and antibiofilm activities; “Host Defence” peptides.
 Modulate Innate & Adaptive Immunity
virulent live MTb strain H37Rv .
After 2 months of infection, treatment
via
New immunomodulatory
peptides
show
IT: 32 g (~1 mg/kg) peptides every 2 days.
broad protection
in Mouse
Model
Infections
Groups of 5 animals
sacrificed after
4 wks
of treatment
Invasive Staph. aureus Mouse
Model and
lung
bacilli
counted.
MDR
-TB Mouse
Model
Cerebral Malaria Mouse Model
100
PLoS One
8:e59119, 2013
p = 0.034
80
Science Transl. Med.
4:135ra64, 2012
70
% Survival
Works vs.
H37Rv &
MDR TB
strains.
d0
4 days (Par 3-6%)
60
Anti-malarial 
Control Peptide
Pyrimethamine +
Chloroquine in
drinking water
50
40
30
20
1018 or
IDR-1
d 4, 5, 6
10
● PBS
■ IDR-1018
Therapeutic IDR-1018 +
Anti-malarial
10 6 PbA
90
IDR 1018:
VRLIVAVRIWRR-NH2
No Treatment
0
0
1
2
3
4
5
6
7
8
9
10 11 12 13
Days post-infection
Science Transl. Med. 4:135ra64, 2012
Also protects vs. E. coli, Salmonella,
Klebsiella, Pseudomonas, MRSA,
In mice
& pigs
VRE, Pox & HSV viruses
Bruno
Rivas
Rogelio
PLoS
One and
7:e39373,
2012 Hernandez Pando, UNA de México
IBD, CF, Sterile inflammation;
Time (days)
LPS/hypoxia-ischemia
 Wound Healing
Lars Steinstraesser, Louis Schofield, Ariel Achtman, Bruno Rivas, Rogelio Hernandez Pando, Carina Mallard
Intranasal delivery (Single dose)
Pseudomonas aeruginosa lung infections
*
(p = 0 .0 1 7 )
T N F a ( p g /m l) in B A L F
6
4
2
80
Harmful
Inflammation 
60
40
20
)
g
)
/k
g
g
m
g
(4
2
(2
0
1
0
0
1
0
1
0
0
2
+
1
0
0
2
2
u
e
s
P
m
m
(1
o
d
o
d
u
e
s
P
/k
/k
g
a
n
o
m
g
r
te
s
a
s
a
n
o
m
)
0
0
W
L o g 1 0 C F U /m l
Protective
Immunity 
100
8
Peptides work by stimulating protective immunity
while suppressing potentially harmful inflammation
Adjuvants and Immunity
Adaptive immunity is antigen-specific, requires gene
rearrangements and thus is slow to develop (days to weeks) and
can discriminate between self and non self.
 Exploited with Vaccines
 Innate immunity fast acting and relatively non-specific.
 Exploited using immune modulators and adjuvants
 But “Innate immunity instructs adaptive immunity” & effectors
TLR4 to NFB interactome
overlap
 Mechanisms still not well understood but:
Adjuvants can either
 Act as a depot (focus)
 Recruit Immune cells
 Activate Immune Cells
Node
Interaction
Need for affordable adjuvants
Adjuvant platform should cost pennies per dose!
Roadblocks to Effective Vaccines
 Need for multiple dosing
(e.g. 3-5 doses for DPT);
Reduces compliance; Delays
protection.
 Maternal interference
 Current adjuvants like Alum
are biased to Th2 responses
 Neonates respond poorly
Vaccination Site
Ivory Coast
 New prime boost regimens
show only partial protection
Volker Gerdts, Lorne Babiuk, Bob Hancock and many others
Triple adjuvant combination gives the potential
for single dose protection vs. pertussis
40,000
30,000
20,000
High Titre
Mixed Th1, Th2
Single Dose
Protective
Pigs, Cattle, Mice
Immunize
Works
in Neonates
Long duration
PTd & PP & CpG-C & HH2
PTd & PP & CpG-C & HH2
Ptd = pertussis toxoid
e.g. Vaccine 31:3148-55, 2013
Also works with RSV, Flu,
Chlamydia prime-boost
PTd & PP & CpG-C
PTd & PP
PTd & CpG-C
PTd & HH2
PTd (pertussis toxoid alone)
10,000
0
Activate Day 0
CpG ODN
Depot
Mouse dose
Day 22
Polyphosphazene; PP
Recruit
IDR
Peptide
0.6 g CpG, 1.2 g
HDP, 0.6 g
Polyphosphazene
Antigen Sparing
Features of the Adjuvant Formulation
Duration of immunity > 2 years (mice)
>10 months in pigs
Comparison to Alum (2 different
doses)
1.0×10 6
IgG2a Antibody Titre
IgG2a Antibody Titre
1.0×10 6
1.0×10 5
1.0×10 4
1.0×10
3
1.0×10 2
1.0×10 1
*
1.0×10 4
*
1.0×10 3
2
4
8
12
16
19
22
26
30
34
38
42
52
61
68
76
85
90
1.0×10 2
Weeks
Bacterial
burden day 2
Protective – Neonatal pigs
2
4
6
8
10
12
14
200000
150000
100000
50000
bo
Tr
ip
le
co
m
ra
ce
l
ua
d
Q
on
tr
ol
0
C
19
Weeks
*
250000
16
Works mucosally at very low doses
*
cfu per total lung lesion
1.0×10 5
Vaccination
OVA plus 0.6 g CpG, 1.2 g
HDP, 0.6 g Polyphosphazene
Acknowledgements
Lab: Ana Nijnik, Neeloffer
Mookherjee, Evan Haney,
Ashley Hilchie, Kelli Wuerth,
Melisssa Elliott, IDR peptide
Team; Bioinformatics Team
Collaborators: Volker Gerdts, VIDO crew, Scott Halperin, Jun
Wang