Presented by:

Anand kumar kushwaha

M.Pharm IInd sem

09321EN017

Department of pharmaceutics ,I.T BHU

Varanasi-221005

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Recent developments in

nanovaccine

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INTRODUCTION

A vaccine is a biological preparation that improvesimmunity to a particular disease.

A vaccine typically contains an agent that resembles adisease-causing microorganism, and is often madefrom weakened or killed forms of the microbe or itstoxins.

The agent stimulates the body's immune system torecognize the agent as foreign, destroy it, and"remember" it, so that the immune system can moreeasily recognize and destroy any of thesemicroorganisms that it later encounters.

WORLD HEALTH ORGANISATION: A REPORT

"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..."

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TYPES OF VACCINE

Vaccine are of three types:

Live

attenuated

vaccine

Killed (Inactivated)

vaccinesToxoids

Exp:

Cholera

Plague

Whopping cough

H.Influenza type b

Exp:

Bacillus-calmette-guerin

Typhoid-ty21a

Poliomyelitis oral

live(OPV)

Exp:

Tetanus(fluid/adsorbed)

Diphtheria(adsorbed)

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NANOVACCINE

Nano is very vast field and it can be applied to anyarea,one of such area is vaccine.

It provide a different routes of administration ofvaccine.

Nanovaccine can be designed, manufactured andintroduced into the human body to improve health,including cellular repairs at the molecular level.

The nanomaterial is so small that it can easily enterthe cell; therefore, nanomaterials can be used in vivoor in vitro for biological applications.

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DELIVERY OF VACCINE:-FOLLOWING ARE THE DIFFERENT WAYTO DELIVER VACCINE AT NANO LEVEL

1.Nanobead: Inert solid bead

Size range 20-200nm.

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.

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Antigen covalently linked to inert nano-beads with a

size of ~50 nm is preferentially taken up by DCs, thus

inducing humoral as well as cell-mediated immune

responses (Scheerlinck et al. 2006)

CONTD…..

*DC;Dendritic cell

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2.POLYMERIC NANOPARTICLES

Biodegradable, biocompatible polymers have

been approved for use in humans.

Poly(D,L-lactide-co-glycolide) (PLG) and

polylactide (PLA).

antigen

adsorbed entrapped

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PLG have been extensively used to encapsulate

antigens.

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.

CONTD….

*PLG: polyglycolide

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3.NANOEMULSION

Size of globule(100-400nm)

Nanoemulsion vaccine does not require refrigeration

and is stable for 6 months.

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.

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4.VIRAL VECTORED VACCINES

Viruses size vary in diameter from 20 nanometres

(nm; 0.0000008 inch) to 250–400 nm.

The immune system quickly respond to viruses, this

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

Modified vaccinia virus ankara (MVA)

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CONTD… Live recombinant viral vector vaccines

are constructed by inserting DNA for the

desired immunogen into a live,

infections but non-pathogenic virus that

elicits a known immune response in

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

vectors..

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CONTD…

Advantages of virally-vectored vaccines include

their ease of production, a good safety profile,

ability to potentiate strong immune responses,

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.

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ADJUVANTS

These substance used in combination with a

specific antigen that produced a more robust

immunity than the antigen alone.

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.

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EXAMPLES

MF59

Monophosphoryl lipid A

Montanide™

Calcium phosphate nanoparticles.

Immunostimulating complexes

Cholesterol-bearing hydrophobized pullulan

nanoparticles (CHP)

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MF59

MF59 is the only nano-sized vaccine adjuvant

approved for human use.

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)

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MONOPHOSPHORYL LIPID A

Monophosphoryl lipid A (MPL®) is an

immunostimulating TLR-4 receptor agonist

composed of detoxified lipopolysaccharide (LPS)

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.

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MONTANIDE™

There are several different types of Montanide™,

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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CONTD….

Emulsions of Montanide™ ISA 51 and 720 are

composed of a metabolizable squalene-based oil with

a mannide monooleate emulsifier.

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

cellular immune responses( A. Saul et.al,1999)

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CALCIUM PHOSPHATE NANOPARTICLES

Calcium phosphate nanoparticles are less than1000nm in diameter.

Nanoparticles can be generated by combining (whilestirring) calcium chloride, sodium phosphate andsodium citrate.

It produces longer duration of response as compare toAl salt.

Phase I study showed that CaP is safe and non-toxicwhen administered subcutaneously.

Vaccines utilizing CaP in preclinical studies includeanthrax, HBV, flu (H5N1 avian and seasonal) andHSV-2.

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*HBV: Hepatitis B virus *HSV: Herpes simplex virus

IMMUNOSTIMULATING COMPLEXES

Another vaccine delivery vehicle with potent

adjuvant activity.

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.

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Quill A: Quillaia saponaria (molina tree)

COMPARISON OF ISCOM WITH CLASSICAL

INFLUENZA VACCINE.

ISCOM induce stronger immune response in

comparison to classical influenza vaccine.

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.

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CONTD…

CHOLESTEROL-BEARING HYDROPHOBIZED

PULLULAN NANOPARTICLES (CHP)

Pullulan is the most popular polysaccharide to whichcholesterol can be Conjugated. It render themolecules amphiphilic.

Such molecules have been shown to self assemblewith and without proteins into 30–40 nm colloidallystable nanoparticles.

Its size and density can be modified by altering thedegree of substitution of cholesterol groups on thepolysaccharide.

A preclinical study in mice showed thatimmunization with a complex of the HER2oncoprotein and CHP induced both humoral and CD8responses.

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*HER: Human epidermal growth factor receptor 2

ADVANTAGES OF NANOVACCINE

Nanovaccine have potential to deliver safe and moreeffective vaccine.

Nanobead covalently coupled with antigen offerdistinct advantages – a low dose of antigen isrequired, efficient processing by antigen-presentingcells and stability during storage.

Encapsulated nanoparticles easily deliver antigen,protects the antigen from degradation and is found tobe effective with a single dose due to slow release ofthe antigen.

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CONTD…

Many of the nanovaccines are non-invasive, delivered

by the oral or nasal route, diffusion patches or

microneedle arrays, thus allowing pain-free delivery

with minimal damage. This is an advantage over

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.

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DISADVANTAGES OF NANOVACCINE

Cost of production.

Nanomaterials can change size, shape but not

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.

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FUTURE PROSPECTS

Carbon nanotubes may be used to deliver vaccine.

Peptide–nano-bead based vaccine approach may be

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

disease ,influenza ,tetanus and HIV based vaccine.

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REFERENCES

1 Tarala D nandedkar, Nanovaccines: recentdevelopments in vaccination, J. Biosci. 34 000–0002009.

2 J. Peek Laura et.al, Nanotechnology in vaccinedelivery, Advanced Drug Delivery Reviews 60 (2008)915–928,

3 J. Bharali Dhruba et.al, Novel nanoparticles for thedelivery of recombinant hepatitis B vaccine,Nanomedicine: Nanotechnology, Biology, andMedicine 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 Biopharmaceutics 55 (2003).

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