Engineering Conferences International ECI Digital Archives

Vaccine Technology IV Proceedings

Spring 5-21-2012 Rational design of a fully synthetic nanoparticle- based for smoking cessatio Takashi Kishimoto Selecta Biosciences

Follow this and additional works at: http://dc.engconfintl.org/vaccine_iv Part of the Biomedical Engineering and Bioengineering Commons

Recommended Citation Takashi Kishimoto, "Rational design of a fully synthetic nanoparticle-based vaccine for smoking cessatio" in "Vaccine Technology IV", B. Buckland, University College London, UK; J. Aunins, Janis Biologics, LLC; P. Alves , ITQB/IBET; K. Jansen, Wyeth Vaccine Research Eds, ECI Symposium Series, (2013). http://dc.engconfintl.org/vaccine_iv/3

This Conference Proceeding is brought to you for free and open access by the Proceedings at ECI Digital Archives. It has been accepted for inclusion in Vaccine Technology IV by an authorized administrator of ECI Digital Archives. For more information, please contact [email protected]. Rational Design of Synthetic

ECI Vaccine Technology Portugal May 2012 Shaping the Immune Response Outline

1) Targeted Particles (tSVP) 2) SEL -068 – Anti - vaccine for

2 The Intersection of Immunology and Nanotechnology

Need for new vaccine technology Robert Langer, ScD Institute Professor • Rapidly mutating pathogens MIT (e.g. HIV, malaria) • Immunosenescence

Omid Farokhzad, MD • Chronic Associate Professor • Cancer Harvard Medical School • Autoimmune diseases and allergies Ulrich von Andrian, • Other chronic diseases MD, PhD Professor (e.g. Alzheimer’s disease, Harvard Medical cardiovascular disease, School smoking cessation)

3 Rational Design of targeted Synthetic Vaccine Particles ( tSVP)

Water Glucose Virus Bacteria Cancer CellA period Tennis . Ball 10 -1 1 10 1 10 2 10 3 10 4 10 5 10 6 10 7 10 8

nanometers

Virus (e.g. influenza virus) tSVP B-Cell Antigen

T-Cell Antigen Adjuvant Biocompatible Matrix

4 Nanoparticles Enable Targeted Delivery of Immunological Instructions to the Lymph Node

•Targeted delivery to DC

Confidential

•Therapeutic Vaccines • Prophylactic Vaccines • Cancer • Infectious diseases • HIV • Therapeutic Vaccines • HCV • Addiction • Other chronic Infections • Cancer

CONFIDENTIAL 5 Self-assembling Manufacturing Process of 4 Synthetic Vaccines

Finished Particles Components Production

PLGA

PEG

B antigen Self- assembly assembly Adjuvant

• Nanoparticle Formation Vessel for • High Batch consistency T antigen 10 g scale (~10,000 doses) • Total batch time <10h (w/o drying and filling)

Advantages of synthetic tSVP production : Successful GMP • Scalability manufacturing • Stable and Reproducible demonstrated to • Fast to the clinic support Ph 1 trial • Cost efficient

6 tSVP-Encapsulated Adjuvant is Targeted to the Draining Lymph Node

Lymph Node IFN-γ Lymph Node IL-12(p40)

30000 140 tSVP [OVA + adjuvant] SVP adjuvant NP-OVA, no adjuvant tSVP [OVA + adjuvant] 120 25000 No adjuvant NP-OVA + admixed adjuvant NP-OVA, no adjuvant Free adjuvant 100 20000 NP-OVA + admixed adjuvant 80

, pg/ml 15000 γ γ γ γ 60

IFN- 10000 40 IL-12 (p40), pg/ml pg/ml (p40), (p40), IL-12 IL-12 5000 20

0 0 0 4 24 48 72 0 4 24 48 72 Time after (hrs) Time after inoculation (hrs)

7 tSVP-Encapsulated Adjuvant Minimizes Systemic Cytokine Production

Plasma TNF-α Plasma IL-6 tSVP [OVA + adjuvant] 14000 tSVP [OVA + adjuvant] 1200 NP-OVA, no aduvant SVP adjuvant NP-OVA, no adjuvant NP-OVA, admixed adjuvant 12000 NP-OVA, admixed adjuvant 1000 No adjuvant Free adjuvant 10000 800 8000 600 6000

400 (pg/mL) IL-6 4000 TNF-alpha (pg/ml) (pg/ml) TNF-alpha TNF-alpha 200 2000

0 0.0 0.5 1.0 1.5 2.0 2.5 3.0 0 1 2 3 4 5 6 Time after inoculation (hrs) Time after (hrs)

•tSVP encapsulation of TLR agonist minimizes systemic cytokine secretion

8 Controlled Adjuvant Release

Adjuvant can be released at a wide range of targeted rates

Percent R-848Adjuvant Release, Release, 37 o 37C C

100

80

60 Release % % Release Release

40 Nanoparticles 62-2

Percent R-848 Release Release Percent Percent R-848 R-848 Nanoparticles 62-4 Percent Percent 20 Nanoparticles 62-6 Nanoparticles 62-8

0 0 50 100 150 200 Time, h

Ideal immune response is correlated with duration of TLR receptor stimulation

9 Universal T-Help Antigen to Boost Response and Enable Universal Vaccines

Algorithm to identify promiscuous MHC class II-binding peptides

Predicted crossreactivity Human PBMCto Selecta's recall Epitope response D 2.5 Allele Allelic variant frequency Peptide D HLA-DRB1*0101 5.4 4.99 2.0 HLA-DRB1*0301 13.7 3.81 HLA -DRB1*0401 4.6 2.02 1.5 HLA-DRB1*0404 3.6 1.79 HLA-DRB1*0405 6.2 4.94 HLA-DRB1*0701 13.5 0.5 1.0 HLA-DRB1*0802 4.9 0.7 % Memory Cells Cells Memory Memory % % HLA-DRB1*0901 6.2 0.36 HLA-DRB1*1101 11.8 3.13 0.5 HLA-DRB1*1302 7.7 1.7 HLA-DRB1*1501 12.2 3.02 0.0

A B C D Epitope Recall response to peptide D observed in 50 of 50 human donors

10 tSVP Induction of Robust Antibody Responses to Wide Range of Antigens

Adjuvant Sparing with Small Molecule Antigen* Viral Peptide Antigen* Protein Antigen*

10X admix adjuvant

TM TM Free tSVP TM w/o tSVP TM Free tSVP tSVP tSVP TM w/o tSVP TM tSVP TM small adjuvant peptide w/o adjuvant w/o w. 1/10 th molecule + alum T cell adjuvant adjuvant + adjuvant antigen +admix adjuvant

* Subcutaneous injection in mice 11 tSVP Induction of In vivo CTL Responses

Fluorescent-labeled target cells

Control peptide Ova peptide pulsed pulsed target cells target cells

tSVP.Ova-immunized Control

18 hrs 18 hrs

Antigen-specific killing of ova- pulsed target cells

12 Efficacy and Memory in E.G7 Tumor Model

Therapeutic Dosing Regimen Survivors re-challenged with tumor 2-3 months later

100 NP-OVA-R848tSVP 100 PBSSaline tSVP -immunized long term survivors 80 80 (n = 8)

60 60

40 40 Non-immunized, tumor-naïve mice

20 (n = 5) Tumor free animals (%) animals free Tumor Tumor free animals20 (%) Tumor-free animals (%) animals Tumor-free

0 0 0 10 20 30 40 50 60 70 6 8 10 12 14 16 18 20 22 Time after tumor inoculation (days) TimeTime after after tumor tumor challenge inoculation (Days) (days)

13 Outline

1) Targeted Synthetic Vaccine Particles (tSVP) 2) SEL -068 – Anti -nicotine vaccine for smoking cessation

14 Smoking Cessation

High prevalence with worldwide smoking population >1B • 46 million adult smokers in USA – 70% desire to quit (CDC) • Smoking costs the US $97.6 billion/yr in direct healthcare costs • 50% of smokers live in BRIC countries High remaining unmet medical need • 12 months abstinence rates for current treatment just over 20% • High relapse risks Smoking cessation is a worldwide priority for health care providers • Fast track status for smoking cessation products in the US • Favorable health economics for smoking cessation products (costs per year of life saved) • High market growth from current $3.8B to over $5.0B in next 5 years

1515 Nicotine Vaccine for Smoking Cessation and Relapse Prevention (SEL-068)

Nicotine vaccine creates nicotine specific that bind free nicotine and prevent it from crossing the blood-brain barrier

Nicotine Universal T help peptide

TLR agonist

1616 Prior Clinical Studies Validate the Vaccine Approach to Smoking Cessation

Nic002 – Cytos (Phase 2)* NicVax - Nabi (Phase 2)** % continuous abstinence (week 8-52) % continuous abstinence (week 8-52) 60% 60%

42% 40% 40% 26% 21% 21% 20% 20% 18%

8% 6%

0% 0%

Two independent studies indicate that an effective vaccine for smoking cessation can be achieved if antibody titers can be increased across a greater percentage of the population

1717 Optimization of an Anti-Nicotine Nanoparticle

1000000

100000 Ab Titer

10000 nicotinenicotine

1000 Anti- -Nicotine-NicotineIg,Ig, GMTGMT max.max.(50%(50% O.D.) O.D.) α α α α

100 Sol NP NP NP NP NP NP NP Nicotine - - Sol - NP Sol NP NP TLR 7/8 agonist - - Sol NP - NP Sol NP MHC II peptide

18 Anti-Nicotine Antibody vs Nicotine AUC

Anti-Nicotine IgG vs Nicotine AUC

19 Persistence of Anti-Nicotine Antibodies

Mice immunized on Days 0, 14, 28, and 180

1000

g/ml) 100 SEL-068 µ µ µ µ No peptide NP 10

1

0.1

Anti-nicotine Ab ( ( Ab Ab Anti-nicotine Anti-nicotine 0.01 0 50 100 150 200 250 300 350 400 450 500 ↑↑↑↑↑↑↑↑↑ ↑↑↑ Days

20 SEL-068: High Antibody Titers Across Mice and Non-Human Primates

Mice Rhesus Monkeys Cynomolgus Monkeys (n=5 per group) (n=4, 1 per group) (n=50, 10 per group) 1000000 1000000 1000000

100000 100000 100000 titer Titer

10000 10000 10000 nicotin nicotin nicotine titer titer nicotine nicotine Nicotiine Nicotiine Anti- Anti- Anti- 1000 1000 1000

100 100 100 e e n n ali osure Sali S p

•R&D and Tox Scale • R&D Scale • Tox scale =P1Nicotine scale pre-ex • d1, d29, d57 • d0, d14, d28 •d0, d28, d56 Nicotine pre-exposure, 8 mg • Antibody titers at d40 • Antibody titers at d70 • Antibody titers at d85

21 High Specificity for Nicotine

Preliminary data Compound % Cross- Compound % Cross- reactivity reactivity

Nicotine 100 Cytisine <1 Cotinine 3.7 Iproniazidphosphate <1

Nornicotine 7.6 Mecamylamine <1 Nicotine -N-Oxide <1 Acetylcholine <1 Norcotinine <1 Choline <1 Cotinine-N-Oxide <1 Niacin <1 Acetylcholine <1 Serotonin <1 BuspironeHCL <1 Dopamine <1 Buproprion HCl <1 γ-Aminobutyric acid (GABA) <1

• Specificity determined by competition ELISA to immobilized poly L-lysine-nicotine • Percent crossreactivity determined as described in Pentel et al., JPET, 2006, 317: 660-666

22 Summary

tSVP Platform • Self-assembling, biocompatible nanoparticles • Directed delivery of TLR agonists to lymph node • Universal T cell help peptide • High antibody titers to poorly immunogenic antigens • Efficient in vivo induction of CTL activity • Antigen-specific immune tolerance SEL-068 Anti-nicotine vaccine • Dose-dependent anti-nicotine response • High titers in mice and two species of non-human primates • Long term persistance of titers • GLP tox study in NHP shows no evidence of systemic toxicities • Successful GMP manufacturing and initiation of Ph 1 23 23 SEL-068 Product Development Timeline

• Selecta begins operations Fall, 2008

• Nicotine program initiated 2009

• >100 formulations made and tested 2009-2010 Start of • Clinical Candidate selected Q4 2010 program to • GLP toxicology study in NHP initiated Feb, 2011 first subject dosed • GLP Tox complete, no systemic findings Aug, 2011 < 3 years • GMP supplies released Aug, 2011

• Phase 1 initiated Nov, 2011

Selecta self-assembling, synthetic nanoparticle platform enables rapid iterative optimization of development candidates and scale up to GMP

24