Download Mucosal vaccines

Nanotechnology
and
mucosal vaccines
Dr. José Crecente Campo
jose·[email protected]
(Mª José Alonso’s group)
Agenda
• Introduction
– What is a vaccine?
– History of vaccination
– Adjuvants
• Mucosal vaccines
– Mucosal immunology
– Nanosystems for mucosal vaccines
– Nasal
• Conclusions
What is a vaccine?
A vaccine typically contains an agent that resembles a disease-causing microorganism and
stimulates the body's immune system to recognize the agent as a threat, destroy it, and keep
a record of it.
www.wikipedia.org
http://www.historyofvaccines.org
History of vaccines
Classic vaccines
 Live attenuated microorganisms
 Inactivated pathogenic agents
Advances in antigen
development
 Recombinant proteins
 Peptides (epitopes)
 Nucleic acids
 Potent immune
response
X Instability
X Adverse side
 Safer
 No need of
adjuvant
effects
X Less immunogenic
 Easier to
produce
X Lower probability of
cross-presentation
 Cheaper
Need potent adjuvants!
Gonzalez-Aramundiz, J. V., Cordeiro, A. S., Csaba, N., de la Fuente, M., & Alonso, M. J. (2012). Nanovaccines: nanocarriers for antigen delivery. Biologie aujourd'hui, 206(4), 249-261.
Cordeiro, A.S.; Alonso, M.J.; De la Fuente, M. Nanoengineering of vaccines using natural polysaccharides. Biotechnology Advances 33 (2015) 1279–1293
Adjuvants
Advances in adjuvant development
Immunostimulatory
molecules
TLR agonists
Bacterial toxins
Particulate delivery systems
Micro
Nano
Saponins
Cytokines
Macromolecules
Nanocarriers
Polysaccharides
Chitosan, dextran,
mannan, beta glucans
 Recognized by
receptors in APCs
 Pathogen-like size and
structure
 Co-encapsulation of
other adjuvant molecules
 Mucosal administration
 Biocompatible /
Biodegradable
 Versatile materials
Cordeiro, A.S.; Alonso, M.J.; De la Fuente, M. Nanoengineering of vaccines using natural polysaccharides. Biotechnology Advances 33 (2015) 1279–1293
Adjuvants
Alum (1930s, Aluminium salts)
Emulsions
Name
Composition
Commercial name
MF59
Squalene O/W
Flu and Focetricia (Seasonal and pandemic influenza)
AS03
Squalene + tocopherol O/W
Pandemrix (Pandemic influenza)
AF03
Squalene O/W
Humenza (Pandemic influenza)
Montanide ISA 51
Mineral oil W/ O
CimaVax EGF* (Non-small lung cancer)
Liposomes
Name
Composition
Commercial name
Inflexal V (seasonal influenza)
Virosome
liposomes
Epaxal (Hepatitis A)
BLP-25 or
emepepimut-S
Biomira liposomal peptide 25
- De Gregorio et al. Vaccines for the twenty-first century society. Nature Reviews Immunology, 2011, 11, 865-872
- Di Pasquale et al. Vaccine Adjuvants: from 1920 to 2015 and Beyond. Vaccines 2015, 3, 320-343
Stimuvax (MUC-1 positive cancer)
Agenda
• Introduction
– What is a vaccine?
– History of vaccination
– Adjuvants
• Mucosal vaccines
– Mucosal immunology
– Nanosystems for mucosal vaccines
– Nasal
• Conclusions
Mucosal vaccines
Systemicmucosal
response
Natural
entrance
Needle-free
vaccination
Mucosal
vaccines
Easier
production
Neutra, M. R., & Kozlowski, P. A. (2006). Mucosal vaccines: the promise and the challenge. Nature Reviews Immunology, 6(2), 148-158.
Mucosal immunology
Large surface
Exposed enviroment
MALT
Inductive/Effector
Lycke, N. (2012). Recent progress in mucosal vaccine development: potential and limitations. Nature Reviews Immunology, 12(8), 592-605.
Mucosal vaccines
Challenge
Challenges
Enzymes
Barriers
Pinto J. Et al., Int. J. Pharm. (2013)
Vicente et al.; J Drug Deliv Sci (2010)
Nanotechnology
Protection
Penetration
Targeting
Ag release
Sustained
Stimulation
Adjuvants
Solution
Nanosystems for mucosal vaccines
SYNTHETIC POLYMERS
Polyesters: PLGA, PLA, PGA
Polyethyleneimine PEI
Acrylic acid based polymers
…
NATURAL POLYMERS
Chitosan
Hyaluronic acid
Alginate, dextran, starch
…
Cordeiro, AS, Alonso, MJ (2015). Recent advances in vaccine delivery. Pharmaceutical Patent Analyst, in press.
Nasal
NALT
-Large surface area
-Reduced enzymatic activity
-Moderately permeable epithelium
-Highly vascularized mucosa
-Absence of first-pass metabolism
-Induce systemic and mucosal immunity
-Easy to administer
-Increase residence time
-Mucoadhesive compounds/help penetration
Potential irritation/irreversible damage longterm
Nasal
PLA
PLA-PEG NPs
TT-I125
Intranasal
Rat nasal mucosa
Tobío et al., 1998
Tobío et al. Stealth PLA-PEG Nanoparticles as Protein Carriers for Nasal Administration. Pharmaceutical Research, Vol. 15, No.2, 1998.
Tobío et al. Improved immunogenicity of a core-coated tetanus toxoid delivery system. Vaccine 18 (2000) 618, 622
Nasal
PLA
PLA-PEG NPs
TT-I125
Intranasal
Tobío et al., 1998
Hypothesis of PEG contribution:
Improved stability in the mucosal fluids
Increased transport across mucosal barriers
Protection/controlled release of the encapsulated antigen
Tobío et al. Stealth PLA-PEG Nanoparticles as Protein Carriers for Nasal Administration. Pharmaceutical Research, Vol. 15, No.2, 1998.
Tobío et al. Improved immunogenicity of a core-coated tetanus toxoid delivery system. Vaccine 18 (2000) 618, 622
Nasal
Chitosan nanoparticles preparation Ionic gelation technique
TPP
Antigen
Chitosan
Chitosan NP
1st report on chitosan nanoparticles: Calvo et al. J. Appl. Pol. Sci, 1997
More than 6000 articles, 100,000 citations
Nasal
2005: “GRAND CHALLENGES IN GLOBAL HEALTH”
GC 1: Create effective single-dose vaccines
GC 3: To develop needle-free delivery systems
for vaccines
Nanostructures as
delivery vehicles for rHBs Antigen
Nasal
MULTIFUNCTIONAL NANOCAPSULES
Antigens:
rHbsAg, influenza HI
Polymeric coating
Polysaccharides,
polyaminoacids
Oils:
squalene, Miglyol, Vitamin-E
Surfactants:
PEG-stearate, Tween 80, TPGS
Nasal
HB antigen
Chitosan
Miglyol
+
Imiquimod
Phospholipids
Vicente, S., Peleteiro, M., Díaz-Freitas, B., Sanchez, A., González-Fernández, Á., & Alonso, M. J. (2013). Co-delivery of viral proteins and a TLR7 agonist from
polysaccharide nanocapsules: a needle-free vaccination strategy. Journal of Controlled Release, 172(3), 773-781.
Nasal
In vitro uptake of chitosan nanocapsules by macrophages
HB antigen
Chitosan
Miglyol
+
Imiquimod
Phospholipids
Cytoskeleton
Nuclei
Nanocapsules
Vicente, S., Peleteiro, M., Díaz-Freitas, B., Sanchez, A., González-Fernández, Á., & Alonso, M. J. (2013). Co-delivery of viral proteins and a TLR7 agonist from
polysaccharide nanocapsules: a needle-free vaccination strategy. Journal of Controlled Release, 172(3), 773-781.
Nasal
Anti-HBsAg IgG (mUI/mL)
Immune response generated by rHBs Ag loaded
nanocarriers following intranasal vs. i.m.
administration
nanocapsules 10µg i.n. (0, 28, 56)
HBsAg-Alum 1µg i.m. (0, 28, 56)
10000
1000
100
Protective
levels
10
1
62
92
122
Time (days)
Sara Vicente et al. PLoS ONE 8(4): e62500, 2013.
152
Nasal
Immune response generated by rHBs Ag loaded
nanocarriers following intranasal administration
S. Vicente et al. Journal of Controlled Release 172 (2013) 773–781
Nasal
Nanocapsules for Single-Shot Vaccination
Anti-HBsAg IgG (mIU/mL)
10000
Day 73
Day 103
Day 133
Day 163
Day 193
1000
100
10
1
NC
10µg (single-dose)
Sara Vicente et al. PLoS ONE 8(4): e62500, 2013.
NC
10µg (0,28)
HBsAg-Alum
10µg (0,28)
Nasal
Elimination of cold chain
Ease of transport and storage
Simple manual resuspension
Thermostable
1
2
3
4
5
6
Completely preserved integrity of rHbsAg after
one year of storage
rHBsAg
25ºC
25ºC
4ºC
HIV-vaccine (NIH project)
Formulation 1
Evaluation
Group 1: prime rVSVpcs, boost NPs
Group 2: prime NPs, boost NPs
Cocktail with 12 peptides NPs
Previous results
Hypothesis
Vaccine targets
1)Protease cleave sites
12 peptides
2)Vital proteins for replication
2 proteins (2 plasmids)
Natural immunity to HIV virus
Formulation 2
Cocktail with 2 plasmids NPs
Non published results. Jorge Pinto et al.
Intranasal administration for NPs
Group 3: prime rVSV prot1prot2, boost DNA-NPs
HIV vaccine
Formulation 1
Chitosan NPs with 1 peptide
Pilot Plant
Microfluidics
- Scale up (small batches)
- GMP production
- Hihgly advanced
characterization techniques
NANOVAC Team
COLLABORATORS
Prof. África González (UVI)
Dr. Belén Díaz-Freitas (UVI)
Mercedes Peleteiro (UVI)
Dr. Ma Luo (U. Manitoba)