Development of Advanced Adjuvants Development of Advanced Adjuvants and Immune Modulatorsand Immune Modulators

R.E.W. (Bob) Hancock,Centre for Microbial Diseases and

Immunity Research,University of British Columbia

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Therapeutic IDR-1018 + Anti-malarial

Anti-malarial Control Peptide

No Treatment

10 6 PbA

d 04 days (Par 3-6%)

1018 or IDR-1d 4, 5, 6

Pyrimethamine + Chloroquine indrinking water

p = 0.034

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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 anti-biofilm activities; “Host Defence” peptides.

Modulate Innate & Adaptive Immunity

Cationic PeptidesCationic PeptidesBirds do it, bees do it, even educated fleas do it.... Cole Porter

New immunomodulatory peptides show New immunomodulatory peptides show broad protection in Mouse Model Infectionsbroad protection in Mouse Model Infections

Lars Steinstraesser, Louis Schofield, Ariel Achtman, Bruno Rivas, Rogelio Hernandez Pando, Carina Mallard

Also protects vs. E. coli, Salmonella, Klebsiella, Pseudomonas, MRSA,

VRE, Pox & HSV viruses

IBD, CF, Sterile inflammation; LPS/hypoxia-ischemia

Wound Healing

IDR 1018: IDR 1018: VRLIVAVRIWRR-NHVRLIVAVRIWRR-NH22

Balb/c mice infected IT with high dose 106

virulent live MTb strain H37Rv .

After 2 months of infection, treatment via IT: 32 g (~1 mg/kg) peptides every 2 days.

Groups of 5 animals sacrificed after 4 wks of treatment and lung bacilli counted.

Works vs. H37Rv &MDR TBstrains.

Bruno Rivas and Rogelio Hernandez Pando, UNA de México

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Therapeutic IDR-1018 + Anti-malarial

Anti-malarial Control Peptide

No Treatment

10 6 PbA

d 04 days (Par 3-6%)

1018 or IDR-1d 4, 5, 6

Pyrimethamine + Chloroquine indrinking water

p = 0.034

MDR -TB Mouse Model Cerebral Malaria Mouse ModelInvasive Staph. aureus Mouse Model

Science Transl. Med. 4:135ra64, 2012

Science Transl. Med. 4:135ra64, 2012

PLoS One 8:e59119, 2013

● PBS■ IDR-1018

In mice& pigs

Time (days)

PLoS One 7:e39373, 2012

Intranasal delivery (Single dose)Intranasal delivery (Single dose)Pseudomonas aeruginosaPseudomonas aeruginosa lung infections lung infections

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1002

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Protective Protective Immunity Immunity

Harmful Harmful Inflammation Inflammation

Peptides work by stimulating protective immunity while suppressing potentially harmful inflammation

Adjuvants and ImmunityAdjuvants and ImmunityAdaptive 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

overlap

Mechanisms still not well understood but:

Adjuvants can either Act as a depot (focus) Recruit Immune cells Activate Immune Cells

Node

Interaction

TLR4 to NFTLR4 to NFB interactomeB interactome

Need for affordable adjuvantsNeed for affordable adjuvants

Adjuvant platform should cost pennies per dose!

Vaccination Site Vaccination Site Ivory CoastIvory Coast

Roadblocks to Effective VaccinesRoadblocks to Effective Vaccines

Volker Gerdts, Lorne Babiuk, Bob Hancock and many others

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

New prime boost regimens show only partial protection

Triple adjuvant combination gives the potential Triple adjuvant combination gives the potential for single dose protection vs. pertussisfor single dose protection vs. pertussis

PTd (pertussis toxoid alone)PTd & HH2

PTd & PPPTd & PP & CpG-C

PTd & CpG-C

PTd & PP & CpG-C & HH2PTd & PP & CpG-C & HH2

PTd & PP & CpG-C & HH2PTd & PP & CpG-C & HH2

CpG ODNIDR

PeptidePolyphosphazene; PP

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Immunize

Ptd = pertussis toxoide.g. Vaccine 31:3148-55, 2013

Also works with RSV, Flu, Chlamydia prime-boost

High TitreMixed Th1, Th2

Single DoseProtective

Pigs, Cattle, MiceWorks in Neonates

Long duration

ActivateActivate DepotDepotRecruitRecruit

Mouse doseMouse dose0.6 0.6 g CpG, 1.2 g CpG, 1.2 g g

HDP, 0.6 HDP, 0.6 g g PolyphosphazenePolyphosphazene

Antigen SparingAntigen Sparing

Duration of immunity > 2 years Duration of immunity > 2 years (mice)(mice)

>10 months in pigs>10 months in pigs

Comparison to Alum (2 different Comparison to Alum (2 different doses)doses)

2 4 6 8 10 12 14 16 191.0×102

1.0×103

1.0×104

1.0×105

1.0×106

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Weeks

IgG

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Weeks

IgG

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Bacterial burden day 2

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Features of the Adjuvant FormulationFeatures of the Adjuvant Formulation

Protective – Neonatal pigsProtective – Neonatal pigs Works mucosally at very low dosesWorks mucosally at very low doses

VaccinationOVA plus 0.6 g CpG, 1.2 g HDP, 0.6 g Polyphosphazene

AcknowledgementsAcknowledgementsLab: Ana Nijnik, Neeloffer Lab: Ana Nijnik, Neeloffer Mookherjee, Evan Haney, Mookherjee, Evan Haney,

Ashley Hilchie, Kelli Wuerth, Ashley Hilchie, Kelli Wuerth, Melisssa Elliott, IDR peptide Melisssa Elliott, IDR peptide Team; Bioinformatics Team Team; Bioinformatics Team

Collaborators: Volker Gerdts, VIDO crew, Scott Halperin, Jun Collaborators: Volker Gerdts, VIDO crew, Scott Halperin, Jun WangWang