1. Recent developments in
nanovaccine
4-May-10
DEPT OF PHARMACEUTICS I.T BHU
Presented by:
Anand kumar kushwaha
M.Pharm IInd sem
09321EN017
Department of pharmaceutics
,I.T BHU
Varanasi-221005
1

2. INTRODUCTION
4-May-10
 A vaccine is a biological preparation that improves
immunity to a particular disease.
DEPT OF PHARMACEUTICS I.T BHU
 A vaccine typically contains an agent that resembles a
disease-causing microorganism, and is often made
from weakened or killed forms of the microbe or its
toxins.
 The agent stimulates the body's immune system to
recognize the agent as foreign, destroy it, and
"remember" it, so that the immune system can more
easily recognize and destroy any of these
microorganisms that it later encounters. 2

3. WORLD HEALTH ORGANISATION: A REPORT
4-May-10
DEPT OF PHARMACEUTICS I.T BHU
"Nearly nine million children under 14 years of
age die every year from infectious disease. And at
least a third of them could be saved if existing
vaccines were more widely used, but the rest only
if
suitable new vaccines were developed..."
3

4. TYPES OF VACCINE
Vaccine are of three types:
4-May-10

DEPT OF PHARMACEUTICS I.T BHU
Killed (Inactivated)
vaccines
Live Toxoids
attenuated
Exp:
Cholera
vaccine Exp:
Plague Tetanus(fluid/adsorbed)
Exp: Diphtheria(adsorbed)
Whopping cough
Bacillus-calmette-guerin
H.Influenza type b
Typhoid-ty21a
Poliomyelitis oral 4
live(OPV)

5. NANOVACCINE
Nano is very vast field and it can be applied to any
4-May-10

area,one of such area is vaccine.
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 It provide a different routes of administration of
vaccine.
 Nanovaccine can be designed, manufactured and
introduced into the human body to improve health,
including cellular repairs at the molecular level.
 The nanomaterial is so small that it can easily enter
the cell; therefore, nanomaterials can be used in vivo
or in vitro for biological applications. 5

6. DELIVERY OF VACCINE:-FOLLOWING ARE THE DIFFERENT WAY
TO DELIVER VACCINE AT NANO LEVEL
4-May-10
1.Nanobead: Inert solid bead
Size range 20-200nm.
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 When antigens are adsorbed on the surface of bead
it has been shown to stimulate CD8 -T cell response.
 The size of the bead play a major role in eliciting a
combined response of humoral and cell-mediated
immunity.
6

7. CONTD…..
 Antigen covalently linked to inert nano-beads with a
size of ~50 nm is preferentially taken up by DCs, thus
4-May-10
inducing humoral as well as cell-mediated immune
responses (Scheerlinck et al. 2006)
DEPT OF PHARMACEUTICS I.T BHU
7
*DC;Dendritic cell

8. 2.POLYMERIC NANOPARTICLES
4-May-10
 Biodegradable, biocompatible polymers have
been approved for use in humans.
DEPT OF PHARMACEUTICS I.T BHU
 Poly(D,L-lactide-co-glycolide) (PLG) and
polylactide (PLA).
antigen
adsorbed entrapped
8

9. CONTD….
 PLG have been extensively used to encapsulate
4-May-10
antigens.
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 PLG forms lactic and glycolic acids, After hydrolysis
of α-hydroxyl acids, yielding small spherical
polymeric particles 1–100 nm in size.
 Adsorbed antigen offer improved stability and
activity over encapsulated antigen by avoiding
exposure to organic solvents used during formulation
and acidic pH conditions caused by degradation of
the polymer. 9
*PLG: polyglycolide

10. 3.NANOEMULSION
Size of globule(100-400nm)
4-May-10

 Nanoemulsion vaccine does not require refrigeration
and is stable for 6 months.
DEPT OF PHARMACEUTICS I.T BHU
 Nanoemulsion is non-toxic, pain free and avoids the
risk of spreading needle-borne infections.(Makidon et
al. 2008)
 Nanoemulsion of hepatitis B antigen, has been
reported to be a safe and effective hepatitis B vaccine.
10

11. 4.VIRAL VECTORED VACCINES
 Viruses size vary in diameter from 20 nanometres
4-May-10
(nm; 0.0000008 inch) to 250–400 nm.
 The immune system quickly respond to viruses, this
DEPT OF PHARMACEUTICS I.T BHU
would seem to be an ideal way to deliver an antigen.
 It consist of a non-replicating virus that contains
some defined genetic material from the pathogen to
which immunity is desired. Such vaccines are also
commonly referred to as live recombinant vaccines.
 Viral vectors are:
Adenovirus
Canary pox virus
Yellow fever viruses 11
Modified vaccinia virus ankara (MVA)

12. CONTD…
 Live recombinant viral vector vaccines
are constructed by inserting DNA for the
desired immunogen into a live,
4-May-10
infections but non-pathogenic virus that
elicits a known immune response in
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humans.
 The most common viral vector is vaccinia
first detailed by Moss in 1987.
Based on vaccinia's success, there are
now more than 20 different RNA and
DNA viruses being tested
for their applications as Vaccine 12
vectors..

13. CONTD…
4-May-10
 Advantages of virally-vectored vaccines include
their ease of production, a good safety profile,
ability to potentiate strong immune responses,
DEPT OF PHARMACEUTICS I.T BHU
potential for nasal or epicutaneous delivery and
mucosal immunization.
 A recent phase I clinical trial of an adenovirus-
vectored flu vaccine administered intranasally
and epicutaneously was found to elicit high
serum antibody titers with a good safety profile.
13

14. ADJUVANTS
4-May-10
 These substance used in combination with a
specific antigen that produced a more robust
immunity than the antigen alone.
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 It improve the immune response to vaccine
antigen by different way:
Increasing immunogenicity of weak antigen.
Enhancing the speed and duration of the
immune response.
Modulating antibody avidity,specifity,isotope
or subclass distribution. 14

15. EXAMPLES
4-May-10
MF59
Monophosphoryl lipid A
DEPT OF PHARMACEUTICS I.T BHU
Montanide™
Calcium phosphate nanoparticles.
Immunostimulating complexes
Cholesterol-bearing hydrophobized pullulan
nanoparticles (CHP)
15

16. MF59
4-May-10
 MF59 is the only nano-sized vaccine adjuvant
approved for human use.
DEPT OF PHARMACEUTICS I.T BHU
 MF59 is an oil-in-water emulsion ( ≤ 250 nm
droplets)
 Its showed a 34-fold increase in antibody titers
when immunized with glycoprotein D of herpes
simplex virus (HSV) in guinea pigs.
 The mechanism of adjuvanticity of MF59 is believed
to be through direct stimulation of cytokine
production (J.K. Simon et.al)
16

17. MONOPHOSPHORYL LIPID A
4-May-10
 Monophosphoryl lipid A (MPL®) is an
immunostimulating TLR-4 receptor agonist
composed of detoxified lipopolysaccharide (LPS)
DEPT OF PHARMACEUTICS I.T BHU
from Salmonella minnesota R595.
 It is a versatile vaccine adjuvant that may be
included in aqueous formulations or in an oil-in-
water emulsion for a more dynamic response.
17

18. MONTANIDE™
4-May-10
 There are several different types of Montanide™,
DEPT OF PHARMACEUTICS I.T BHU
including ISA 50V, 51, 206 and 720.
 ISA 50V, 51 and 720 are water-in-oil emulsions
while ISA 206 is a water-in-oil-in-water
emulsion.
 ISA 206 and 50V only used in veterinary vaccine
formulations.
 ISA 51 & ISA 720 are under investigation for use
in humans.
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19. CONTD….
 Emulsions of Montanide™ ISA 51 and 720 are
4-May-10
composed of a metabolizable squalene-based oil with
a mannide monooleate emulsifier.
DEPT OF PHARMACEUTICS I.T BHU
 The immune enhancement produced by the
Montanide™ emulsions is believed to be due to the
formation of a depot at the site of injection. (A.P.
Miles et.al, 2005)
 The emulsion vaccines against malaria, HIV and
various cancer have been in phase I and/or II clinical
trials.
 A phase I trial of a trivalent malaria vaccine
containing ISA 720 induced both humoral and
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cellular immune responses( A. Saul et.al,1999)

20. CALCIUM PHOSPHATE NANOPARTICLES
4-May-10
 Calcium phosphate nanoparticles are less than
1000nm in diameter.
DEPT OF PHARMACEUTICS I.T BHU
 Nanoparticles can be generated by combining (while
stirring) calcium chloride, sodium phosphate and
sodium citrate.
 It produces longer duration of response as compare to
Al salt.
 Phase I study showed that CaP is safe and non-toxic
when administered subcutaneously.
 Vaccines utilizing CaP in preclinical studies include
anthrax, HBV, flu (H5N1 avian and seasonal) and
HSV-2. 20
*HBV: Hepatitis B virus *HSV: Herpes simplex virus

21. IMMUNOSTIMULATING COMPLEXES
4-May-10
 Another vaccine delivery vehicle with potent
adjuvant activity.
DEPT OF PHARMACEUTICS I.T BHU
 Produced by combining a protein antigen,
cholesterol, phospholipid and the saponin
adjuvant Quil A.
 These are ~40 nm cage-like particles.
 The matrix that is formed traps the protein
antigens.
21
Quill A: Quillaia saponaria (molina tree)

22. CONTD…
COMPARISON OF ISCOM WITH CLASSICAL
INFLUENZA VACCINE.
4-May-10
 ISCOM induce stronger immune response in
comparison to classical influenza vaccine.
DEPT OF PHARMACEUTICS I.T BHU
 ISCOM based flu vaccine showed that virus-
specific CTL memory was achieved in 50–60% of
the patients while in case of classical influenza it
is only 5%.
 Intranasally administration of ISCOM flu
vaccine showed strong mucosal (IgG and IgA)
responses as well as systemic and CTL responses.
 Oral vaccine of ISCOM also effective but requires
high and frequent dosing. 22

23. CHOLESTEROL-BEARING HYDROPHOBIZED
PULLULAN NANOPARTICLES (CHP)
4-May-10
 Pullulan is the most popular polysaccharide to which
cholesterol can be Conjugated. It render the
molecules amphiphilic.
DEPT OF PHARMACEUTICS I.T BHU
 Such molecules have been shown to self assemble
with and without proteins into 30–40 nm colloidally
stable nanoparticles.
 Its size and density can be modified by altering the
degree of substitution of cholesterol groups on the
polysaccharide.
A preclinical study in mice showed that
immunization with a complex of the HER2
oncoprotein and CHP induced both humoral and CD8
responses. 23
*HER: Human epidermal growth factor receptor 2

24. ADVANTAGES OF NANOVACCINE
4-May-10
 Nanovaccine have potential to deliver safe and more
effective vaccine.
DEPT OF PHARMACEUTICS I.T BHU
 Nanobead covalently coupled with antigen offer
distinct advantages – a low dose of antigen is
required, efficient processing by antigen-presenting
cells and stability during storage.
 Encapsulated nanoparticles easily deliver antigen,
protects the antigen from degradation and is found to
be effective with a single dose due to slow release of
the antigen.
24

25. CONTD…
 Many of the nanovaccines are non-invasive, delivered
4-May-10
by the oral or nasal route, diffusion patches or
microneedle arrays, thus allowing pain-free delivery
with minimal damage. This is an advantage over
DEPT OF PHARMACEUTICS I.T BHU
conventional vaccines,which are usually multi-
injection, multi-dose delivery systems.
 The nanoemulsion preparation of hepatitis B antigen
found to be tolerable and effective and does not
require refrigeration and it is effective for a month at
25ºc and for 6 week at 40ºc,therefore it facilitate its
final distribution in small areas/villages of developing
countries. 25

26. DISADVANTAGES OF NANOVACCINE
4-May-10
 Cost of production.
 Nanomaterials can change size, shape but not
DEPT OF PHARMACEUTICS I.T BHU
composition, which may change their toxicity.
 Small nanoparticles are cleared quickly from the
body, large counterparts may accumulate in vital
organs causing toxic problems.
 Reproducibility of formulation during manufacturing
is one of the major hurdles in the use of
nanoparticles as vaccines.
26

27. FUTURE PROSPECTS
4-May-10
 Carbon nanotubes may be used to deliver vaccine.
 Peptide–nano-bead based vaccine approach may be
DEPT OF PHARMACEUTICS I.T BHU
beneficial, especially for highly variable pathogens
such as FMDV(foot and mouth disease virus).
 Nanoemulsion may deliver smallpox, influenza,
anthrax and HIV vaccines.
 Nanoemulsion against GP 120, one of the major
binding proteins, may induce mucosal and cellular
immunity, and neutralize antibody to various
isolates of HIV.
 Adenovirus may deliver vaccine for Alzheimer's
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disease ,influenza ,tetanus and HIV based vaccine.

28. REFERENCES
4-May-10
1 Tarala D nandedkar, Nanovaccines: recent
developments in vaccination, J. Biosci. 34 000–000
2009.
DEPT OF PHARMACEUTICS I.T BHU
2 J. Peek Laura et.al, Nanotechnology in vaccine
delivery, Advanced Drug Delivery Reviews 60 (2008)
915–928,
3 J. Bharali Dhruba et.al, Novel nanoparticles for the
delivery of recombinant hepatitis B vaccine,
Nanomedicine: Nanotechnology, Biology, and
Medicine 4 (2008) 311–317.
4 Cui Zhengrong et.al, The effect of co-administration of
adjuvants with a nanoparticle-based genetic vaccine
delivery system on the resulting immune responses,
European Journal of Pharmaceutics and 28
Biopharmaceutics 55 (2003).

29. 4-May-10 DEPT OF PHARMACEUTICS I.T BHU
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