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By
                    Dr. Shreeraj Shah
                Associate Professor,
Dept. of Pharmaceutical Technology,
          L.J. Institute of Pharmacy,
                          Ahmedabad
                                  1
Content
Introduction
General characteristics of Niosomes
Advantage of Niosomes
Disadvantage of Niosomes
Structure of Niosomes
Contrast and similarity between Niosomes and Liposomes
Methods of Preparation
Factors affecting the physicochemical properties of Niosomes
Types of Niosomes
Characterization of Niosomes
Therapeutic Applications
Marketed products
Future Prospects
Conclusion
                                                                2
References
Introduction
• Development of new drug, improving safety and efficacy of existing drugs is
  difficult, expensive and time consuming
• At present, no available drug delivery system behaves ideally achieving all
  the lofty goals
• Encapsulation of the drug in vesicular structures is one of the promising
  system
• such as liposomes, niosomes, transferosomes, ethosomes and pharmacosomes
  etc
• It delivers drug directly to the site of action, leading to reduction of drug
  toxicity with no adverse effects                                         3
•Vesicular drug delivery reduces the cost of therapy by
improving bioavailability of medication and also solves the
problems of drug insolubility, instability and rapid degradation

•Niosomes are a novel drug delivery system, in which the
medication is encapsulated in a vesicle, composed of a bilayer
of non-ionic surface active agents and hence the name
niosomes

•The niosomes are very small, and microscopic in size. Their
size lies in the nanometric scale

                                                            4
Definition
• Niosomes are synthetic microscopic vesicles
 consisting of an aqueous core enclosed in a bi
 layer consisting of cholesterol and one or more
 nonionic surfactants
• Vesicles are prepared from self assembly of
 hydrated non ionic surfactants molecules
                                              5
General Characteristics of Niosome
•Biocompatible, biodegradable, non-toxic, non immunogenic and
non-carcinogenic
•The ability of nonionic surfactant to form bilayer vesicles is
dependant on the HLB value of the surfactant, the chemical structure
of the components and the critical packing parameter
•Niosomes can be characterized by their size distribution studies
•   High resistance to hydrolytic degradation
•The properties of niosome depends both on composition of the
bilayer & on method of their production
                                                                    6
Advantages of Niosomes
 Targeted drug delivery can be achieved using niosomes the drug is delivered directly to
  the body part where the therapeutic effect is required
 Reduced dose is required to achieve the desired effect
 Subsequent decrease in the side effects
 The therapeutic efficacy of the drugs is improved by reducing the clearance rate, targeting
  to the specific site and by protecting the encapsulated drug
 Niosomes are amphiphillic i.e. both hydrophilic and lipophillic in nature and can
  accommodate a large number of drugs with a wide range of solubilities
 Improve the oral bioavailability of poorly soluble drugs
 Enhance the skin permeability of drugs when applied topically
 provide advantage of usage through various routes viz. oral, parentral, topical, ocular etc.
 The bilayers of the niosomes protect the enclosed Active pharmaceutical ingredient from
  the various factors present both inside and outside the body
 The surfactants used and also the prepared niosomes are biodegradable, biocompatible
  and non-immunogenic
 They are osmotically active and stable

                                                                                          7
Disadvantages of Niosome
•Aqueous suspension of niosome may exhibit fusion, aggregation
leaching or hydrolysis of entrapped drug, thus limiting the shelf life
of niosome dispersion.
• Time consuming
• Requires specialized equipment
• Inefficient drug loading




                                                                    8
Structure of Niosomes
•Niosomes are microscopic lamellar structures
•Basic structural components are
          Non ionic surfactant
          Cholesterol
          Charge inducing molecule
•A number of non-ionic surfactants used are:
   polyglycerol alkyl ether, glucosyl dialkyl ethers, crown ethers, ester
linked surfactants, polyoxyethylene alkyl ether and a series of spans and
tweens

                                                                        9
Non-ionic Surfactants
The non-ionic surfactants orient themselves in bilayer lattices

  where the polar or hydrophobic heads align facing aqueous
  bulk (media) while the hydrophobic head or hydrocarbon
  segments align in such a way that the interaction with the
  aqueous media would be minimized.
 To attain thermodynamic stability, every bilayer folds over

  itself as continuous membrane i.e. forms vesicles so that
  hydrocarbon/water interface remains no more exposed.
                                                             10
Mainly following types of non-ionic surfactants are used for the formation of niosomes:
 Alkyl Ethers: L’Oreal described some surfactants for the preparation of niosomes
  containing drugs/chemicals as
    1) Surfactant-I (molecular weight (MW 473)) is C16 monoalkyl glycerol ether with
      average of three glycerol units.
    2) Surfactant-II (MW 972) is diglycerol ether with average of the seven glycerol units.
    3) Surfactant III (MW 393) is ester linked surfactant.
  Other than alkyl glycerol, alkyl glycosides and alkyl ethers bearing polyhydroxyl head
  groups are also used in formulation of niosomes.
 Alkyl Esters: Sorbitan esters are most preferred surfactant used for the preparation of
  niosomes amongst this category of surfactants. Vesicles prepared by the polyoxyethylene
  sorbitan monolaurate are relatively soluble than other surfactant vesicles. For example
  polyoxyethylene (polysorbate 60) has been utilized for encapsulation of diclofenac
  sodium. A mixture of polyoxyethylene-10-stearyl ether : glyceryl laurate : cholesterol (27 :
  15 : 57) has been used in transdermal delivery of cyclosporine-A.
 Alkyl Amides: Alkyl amide (e.g. galactosides and glucosides) have been utilized to
  produce niosomal vesicles.
 Fatty Acid and Amino Acid Compounds: Long chain fatty acids and amino acid moieties
  have also been used in some niosomes preparation.                                       11
Diff. types of Non-ionic surfactants




                                       12
Cholesterol
Steroids are important components of the cell membrane and their
 presence in membrane affect the bilayer fluidity and permeability.
Cholesterol is a steroid derivative, which is mainly used for the
 formulation of niosomes. Although it may not show any role in the
 formation of bilayer, its importance in formation of niosomes and
 manipulation of layer characteristics can not be discarded.
In general, incorporation of cholesterol affects properties of niosomes
 like membrane permeability, rigidity, encapsulation efficiency, ease of
 rehydration of freeze dried niosomes and their toxicity.
It prevents the vesicle aggregation by the inclusion of molecules that
 stabilize the system against the formation of aggregates by repulsive
 steric or electrostatic forces that leads to the transition from the gel to
 the liquid phase in niosome systems. As a result of this, the niosome
 becomes less leaky in nature.                                          13
Charge inducing molecule
Some charged molecules are added to niosomes to increase stability

  of niosomes by electrostatic repulsion which prevents aggregation
  and coalescence. The negatively charged molecules used are
  diacetyl phosphate (DCP) and phosphotidic acid. Similarly,
  stearylamine (STR) and stearyl pyridinium chloride are the well
  known    positively   charged    molecules    used   in   niosomal
  preparations.
Only 2.5-5 mol % concentration of charged molecules is tolerable

  because high concentration can inhibit the niosome formation
                                                                 14
15
Contrast of Niosomes Vs liposomes
         Niosome                      Liposome

    1. Less Expensive             1. More expensive
   2. Chemically Stable         2. Chemically unstable
3. Niosomes are prepared      3. liposomes are prepared
   from uncharged single-           from double-chain
      chain surfactant                phospholipids
  4. They do not require       4. They require special
     special storage and          storage, handling &
          handling                  purity of natural
                                phospholipid is variable.
5. Non ionic drugs carriers   5. The ionic drugs carriers
           are safer              are relatively toxic &
                                        unsuitable
                                                            16
Similarity of niosomes and liposomes
• However Niosomes are similar to liposomes in functionality.
• Niosomes also increase the bioavailability of the drug and
  reduce the clearance like liposomes.
• Niosomes can also be used for targeted drug delivery, similar
  to liposomes.
• As with liposomes, the properties of the niosomes depend
  both- on the composition of the bilayer, and the method of
  production used.

                                                                  17
Method of Preparation
1. Ether Injection (LUV)             Based on the vesicle size, niosomes
                                      can be divided into three groups.
2. Hand Shaking Method (MLV)         These are small unilamellar vesicles
                                         (SUV, size=0.025-0.05 μm),
3. The “Bubble” Method                  multilamellar vesicles (MLV,
                                          size=>0.05 μm), and large
4. Reverse Phase Evaporation (LUV)        unilamellar vesicles (LUV,
                                               size=>0.10 μm).
5. Sonication (SUV)
6. Multiple membrane extrusion method
7. Trans Membrane pH Gradient Drug Uptake Process (remote
   Loading) (MLV)
8. Microfluidization method (SUV)
9. Formation of Niosomes From Proniosomes                            18
General steps of Niosome preparation

• Hydration of mixture of surfactant/lipid at elevated
 temperature
• Sizing of niosomes

• Removal of unentrapped material from vesicles




                                                         19
Common stages of all methods of preparation of Niosomes
                Cholesterol + Non ionic surfactant

                               Dissolve in organic solvent

                   Solution in organic solvent


                                Drying

                         Thin film

                                Dispersion (Hydration)


                      Niosome suspension

                                                             20
1. Ether injection method

•   This method is based on slow injection of surfactant : cholesterol
    solution in ether through 14 gauge needle into a preheated aqueous
    phase maintained at 600C

•   Vaporization of ether resulting into a formation of ether gradient at
    ether-water interface which leads to formation of single layered
    vesicles

•   Depending upon the conditions used, the diameter of the vesicle
    range from 50 to 1000 nm



                                                                    21
22
2. Hand shaking method
(Thin film hydration technique)

•   Surfactant and cholesterol are dissolved in a volatile organic solvent
    (diethyl ether, chloroform or methanol) in a round bottom flask

•   The organic solvent is removed under vaccum at room temperature
    using rotary evaporator leaving a thin layer of solid mixture
    deposited on the wall of the flask

•   The dried surfactant film can be rehydrated with aqueous phase at
    temperature slightly above the phase transition temperature of the
    surfactant used, with gentle agitation

•   This process forms large multilamellar niosomes                  23
24
3. The “Bubble” Method

•It is novel technique for the one step preparation of liposomes and
niosomes without the use of organic solvents

•The bubbling unit consists of round-bottom flask with three necks
positioned in water bath to control the temperature

•Water-cooled reflux and thermometer is positioned in the first and
second neck and nitrogen supply through the third neck

• Cholesterol and surfactant are dispersed together in this buffer (pH 7.4)
at 70°C, the dispersion mixed for 15 seconds with high shear
homogenizer and immediately afterwards “bubbled at 70°C using
nitrogen gas
                                                                      25
4. Reverse Phase Evaporation Technique (REV)

•Cholesterol and surfactant (1:1) are dissolved in a mixture of ether
and chloroform

•An aqueous phase containing drug is added to this and the resulting
two phases are sonicated at 4-5°C. The clear gel formed is further
sonicated after the addition of a small amount of phosphate buffered
saline (PBS)

•The organic phase is removed at 40°C under low pressure. The
resulting viscous niosome suspension is diluted with PBS and heated
on a water bath at 60°C for 10 min to yield niosomes

                                                                   26
27
5. Sonication
•In this method, an aliquot of drug solution in buffer is added to the
surfactant/cholesterol mixture in a 10-ml glass vial.
•The mixture is probe sonicated at 60°C for 3 minutes using a sonicator.
•The resultant vesicles are of small unilamellar type niosomes.

6. Multiple membrane extrusion method
• In this method, a mixture of surfactant, cholesterol and diacetyl phosphate is prepared
and then solvent is evaporated using rotary vacuum evaporator to leave a thin film. The
film is then hydrated with aqueous drug solution and the suspension thus obtained is
extruded through the polycarbonate membrane (mean pore size 0.1 mm) and then
placed in series up to eight passages to obtain uniform size niosomes.
•Good method of controlling niosome size.                                          28
Sonication method   Extrusion method


                                   29
7. Trans membrane pH gradient (inside acidic) Drug
   Uptake Process (remote Loading)
•Surfactant and cholesterol are dissolved in chloroform
• The solvent is then evaporated under reduced pressure to get a thin film on the
wall of the round bottom flask
•The film is hydrated with 300 mM citric acid (pH 4.0) by vortex mixing.
•The multilamellar vesicles are frozen and thawed 3 times and later sonicated.
 To this niosomal suspension, aqueous solution containing 10 mg/ml of drug
 is added and vortexed
• The pH of the sample is then raised to 7.0-7.2 with 1M disodium phosphate
• This mixture is later heated at 60°C for 10 minutes to give niosomes
                                                                            30
8. Microfluidization Method
In this method two fluidized streams (one containing drug

 and the other surfactant) interact at ultra high velocity, in
 precisely defined micro channels within the interaction
 chamber in such a way that the energy supplied to the
 system remains in the area of niosomes formations. This is
 called submerged jet principle. It results in better
 uniformity, smaller size and reproducibility in the
 formulation of niosomes
                                                           31
9. Formation of Niosomes from Proniosomes
•Another method of producing niosomes is to coat a water-soluble
carrier such as sorbitol with surfactant
• The result of the coating process is a dry formulation. In which each
water-soluble particle is covered with a thin film of dry surfactant. This
preparation is termed “Proniosomes”
•The niosomes are formed by the addition of aqueous phase at T > T m
and brief agitation
                         T = Temperature.
                        Tm = mean phase transition temperature




                                                                     32
Separation of unentrapped drugs
The removal of unentrapped solute from the vesicles can be
accomplished by various techniques, which include:
Gel Filtration
The unentrapped drug is removed by gel filtration of niosomal dispersion through a
Sephadex-G-50 column and elution with phosphate buffered saline or normal saline.

Dialysis
The aqueous niosomal dispersion is dialyzed in a dialysis tubing against phosphate
buffer or normal saline or glucose solution

Centrifugation
The niosomal suspension is centrifuged and the supernatant is separated. The pellet is
washed and then resuspended to obtain a niosomal suspension free from unentrapped
                                                                                   33
drug.
Factors affecting the physicochemical properties of
niosomes
Membrane Additives
  Stability of niosomes can be increased by the number of
    additives into niosomal formulation along with surfactant and
    drugs. The membrane stability, morphology and permeability of
    vesicles are affected by numbers of additives e.g. addition of
    cholesterol in niosomal system increases the rigidity and
    decreases the drugs permeability through the membrane
Temperature of Hydration
  Shape and size of niosome is also influenced by the hydration
    temperature. Assembly of the niosomes vesicles is affected by
    the temperature change of niosomal system. Temperature change
    can also induce the vesicle shape transformation          34
Properties of Drugs
    The drug entrapment in niosomes is affected by molecular weight, chemical
     structure, hydrophilicity, lipophilicity as well as the hydrophilic lipophilic
     balance (HLB) value of the drug. Vesicle size may increase due to entrapment
     of drug. Drug particle interact with the surfactant head groups, which may
     increase charge on polymer and thus cause repulsion of the surfactant bilayer
     which leads to increase in vesicle size.
Amount and Type of Surfactant
    As the HLB value of surfactants like span 85 (HLB 1.8) to span 20 (HLB 8.6)
     increased, the mean size of niosomes also increases proportionally. It is due to
     the fact that surface free energy decreases with increase in hydrophilicity of
     surfactant. Alkyl chain is present in well ordered structure in gel state, while
     in the liquid state the structure of bilayer is more disordered.
    Entrapment efficiency is also affected by phase transition temperature i.e.
     span 60 having higher TC, provide better entrapment efficiency. Entrapment
     efficiency of the niosomes is affected by the HLB value for e.g. niosomes
     have high entrapment efficiency at HLB value 8.6 but HLB value 14 to 17 is
                                                                                 35
     not suitable for niosomes formulation
Effect of the nature of drug on the formation of niosomes




                                                            36
Cholesterol Content and charge
   Hydrodynamic diameter and entrapment efficiency were found to be increased due to
    cholesterol content in the niosomal bilayer. Cholesterol can act by two ways. First, it can
    increase the chain order of liquidated bilayer and second, by decreasing the chain order of
    the gel state bilayer.
   Release rate of drug decreases and rigidity of bilayer increases due to high concentration of
    cholesterol
   The presence of charge tends to increase the interlamellar distance between successive
    bilayers in multilamellar vesicle structure and leads to greater overall entrapped volume

Structure of Surfactants
   The geometry of vesicles to be formed from surfactant is affected by its structure, which is
    related to critical packing parameter (CPP)



                   where V= hydrophobic group volume
                   Ic = the critical hydrophobic group length
                   a0 = the area of hydrophilic head group
    spherical micelles formed if CPP<0.5, inverted micelles is formed if CPP>1.             37
Method of Preparation
    The average size of acyclovir niosomes prepared by hand-shaking process
     was larger (2.7m m) as compared to the average size of niosomes 1.5m m
     prepared by ether injection method which may be attributed to the passage of
     cholesterol and span-80 solution through an orifice into the drug solution.
     Reverse phase evaporation can be used to produce smaller size vesicles.
     Vesicles with smaller size and greater stability can be produced by
     microfluidization method. Niosomes obtained by transmembrane pH gradient
     (inside acidic) drug uptake process showed greater entrapment efficiency and
     better retention of drug
Resistance to Osmotic Stress
    Diameter of niosomal vesicles was found to be decreased when niosomal
     suspension is kept in contact with hypertonic salt solution. There is slow
     release with slight swelling of vesicles, which is due to inhibition eluting
     fluids from vesicles, followed by faster release, which may be due to decrease
     in mechanical strength under osmotic stress

                                                                              38
Types of Niosomes
Proniosomes
  Proniosomes are the niosomal formulation containing carrier and
   surfactant, which requires to be hydrated before being used. The
   hydration results in the formation of aqueous niosome dispersion.
   Proniosomes decreases the aggregation, leaking and fusion problem
   associated with niosomal formulation
Aspasomes
  Combination of acorbyl palmitate, cholesterol and highly charged
   lipid diacetyl phosphate leads to the formation of vesicles called
   aspasomes. Aspasomes are first hydrated with water/aqueous
   solution and then sonicated to obtain the niosomes. Aspasomes can
   be used to increase the transdermal permeation of drugs. Aspasomes
   have also been used to decrease disorder caused by reactive oxygen
   species as it has inherent antioxidant property                39
Vesicles in Water and Oil System (v/w/o)
  It has been reported that the emulsification of an aqueous
    niosomes into an oil phase form vesicle in water in oil emulsion
    (v/w/o). This can be prepared by addition of niosomes
    suspension formulated from mixture of sorbitol monostearate,
    cholesterol and solulan C24 (poly-24-oxyethylene cholesteryl
    ether) to oil phase at 60°C. This results in the formation of
    vesicle in water in oil (v/w/o) emulsion which by cooling to
    room temperature forms vesicle in water in oil gel (v/w/o gel).
    The v/w/o gel thus obtained can entrap proteins/proteinous drugs
    and also protect it from enzymatic degradation after oral
    administration and controlled release. The immunogenic
    properties of v/w/o gel and w/o gel reported that both exhibit
    immunoadjuvant tendency. In this system aqueous niosomes
    (v/w) are emulsified into the oily phase
                                                                40
Niosomes in Carbopol Gel
  Niosomes were prepared using drug, spans and cholesterol. The
    niosomes thus obtained were then incorporated in carbopol-934
    gel (1% w/w) base containing propylene glycol (10% w/w) and
    glycerol (30% w/w). It was observed that the mean flux value
    and diffusion co-efficient were 5 to 7 times lower for niosomal
    gel as compared to plain drug gels. Moreover, carrageenan
    induced paw edema inhibition (i.e. 66.68 ± 5.19%) was higher by
    niosome formulation as compared to plain gel.
Niosomes of Hydroxyl Propyl Methyl Cellulose
  In this type, a base containing 10% glycerin of hydroxy propyl
    methyl cellulose was first prepared and then niosomes were
    incorporated in it. The bioavailability and reduction of paw
    edema induced by carrageenan was found to be higher by this
    niosomal system than the plain formulation of flurbiprofen. 41
Characterization of niosomes
Physical characterization

Chemical characterization

Biological characterization




                               42
Physical characterization
Sr no.   Parameters                     Analytical
                                        methods/Instruments
1.       Vesicles shape & surface       Transmission electron
         morphology                     microscopy,
                                        Freeze-fracture electron
                                        microscopy
2.       Mean vesicle size and size   Dynamic light scattering,
         distribution                 zetasizer,
         (submicron and micron range) Photon correlation
                                      spectroscopy,
                                      laser light scattering, gel
                                      permeation and gel
                                      exclusion

3.       Surface charge                 Free-flow electrophoresis
                                                               43
4.   Phase behavior                  Freeze-fracture electron
                                    microscopy, Differential
                                    scanning colorimetery

5.   Electrical surface potential   Zeta potential
     and surface pH                 measurements & pH
                                    sensitive probes


6.   Entrapment efficiency          Amount entrapped/total
                                    amount added*100




                                                            44
 Chemical characterization
 Sr no   Parameters                   Analytical
                                      methods/Instruments
 1.      Cholesterol concentration    Cholesterol oxidase
                                      assay and HPLC


 2.      Cholesterol auto-oxidation   HPLC and TLC




 3.      Osmolarity                   Osmometer



                                                            45
 Biological characterization
 Sr No.    Parameters       Analytical
                            methods/Instruments
 1.       Sterility         Aerobic or anaerobic cultures



 2.       Pyrogenicity      Limulus Amebocyte Lysate
                            (LAL) test


 3.       Animal toxicity   Monitoring survival rates,
                            histology and pathology

                                                            46
Evaluation Parameters (Summary)
 % Entrapment Efficiency (%EE)
 Size, Shape and Morphology
      Transmission Electron Microscopy (TEM)
      Freeze Fractured Microscopy

      Optical Microscopy Technique

 In Vitro Release Study
 Tissue Distribution/In Vivo Study
 Stability Study
 Number of Lamellae
 Membrane Rigidity
 Vesicular Surface Charge
                                                47
Drugs and routes of administration used in
                    Niosomes
      1. Intravenous Route               Ipromide, Vincristine,Indomethacin,
                                        Colchicines, Rifampicin, Transferrin,
                                        Zidovudine, Cisplantin, amarogentin,
                                           Daunorubicin, AmphotericinB.
     2. Transdermal Route             Flurbiprofen, Piroxicam, Levonorgestrol,
                                        Nimeluside, Estradiol, Ketoconazole,
                                    Enoxacin, DNA loaded noisome, Cyclosporine,
                                             Erythromycin, α interferon.
         3. Oral Route                Vaccine, Polysaccharide Coated noisome,
                                               Cipro floxacin, Insulin.
       4. Oncology Route               Methotrexate, Doxorubicin, adriamycin.
        5. Ocular Route                        Timolol, cyclopentolate
         6. Nasal Route                        Sumatriptan, Influenza
   7.Immunological Adjuvant             Bovine serum albumin, Hemoglobin

8. For treatment of Leishmaniasis                 Stilbogluconat
                                                                            48
Therapeutic applications of Niosomes
The applications of niosomes can be mainly classified into
 three categories

  1) For Controlled Release of Drugs

  2) To Improve the Stability and Physical

      Properties of the Drugs

  3) For Targeting and Retention of Drug in

      Blood Circulation
                                                        49
1)To Prolong the Release Rate of Drugs
1.1
     For Controlled Release
  The release rate of drugs like withaferin and gliclazide from the
    niosomes was found slower as compared to other dosage forms
1.2 In Ophthalmic Drug Delivery
  Experimental results of the water soluble antibiotic gentamicin
   sulphate showed a substantial change in the release rate. Beside
   this, the percent entrapment efficiency of gentamicin sulphate
   was altered when administered as niosomes. Also, as compared
   to normal drug solution, niosomes of drug show slow release
  Niosomal formulation containing timolol maleate (0.25%)
   prepared by chitosan coating exhibited more effect on intra
   ocular tension with fewer side effects as compared to the
   marketed formulation.
                                                                 50
2) To Improve the Stability and Physical
 Properties of the Drugs
2.1 To Increase Oral Bioavailability
    With the formulation of niosomes, the oral bioavailability of the acyclovir as
     well as griseofulvin was increased as compared to the drug alone. Similarly,
     the absorptivity of poorly absorbed peptide and ergot alkaloid can be
     increased by the administration in the bile duct of rats when administered as
     micellar solution together with the POE-24-cholesteryl ester
2.2 For Improvement of Stability of Peptide Drugs
    8-arginin vasopressin, 9-glycinamide-ω
    The in vitro release of insulin from niosomes formulated by span 40 and span
     60 in simulated intestinal fluid was lower than the niosomes formulated by
     span 20 and span 80.
    Niosomes prepared by the span 60 has high resistance against proteolytic
     enzyme and exhibit good stability in storage temperature and in presence of
     sodium deoxycholate                                                    51
2.3 To Promote Transdermal Delivery of Drugs
    Many drugs such as lidocaine, estradiol, cyclosporine etc. are used for topical
     and transdermal drug delivery system by formulating them as niosomes
    The niosomes of natural compound, ammonium glycyrrhizinate were
     formulated for effective anti-inflammatory activity using new non-ionic
     surfactant, bola surfactat-span 80-cholesterol (2 : 3 : 1 ratio).
    Experimental study showed that the bola niosomes were able to promote the
     intracellular uptake of ammonium glycerrhizinic acid
2.4 As a Tool for Improvement of Stability of Immunological Products
    Important tool for immunological selectivity, low toxicity and more stability
     of the incorporated active moiety
2.5 To Improve Anti-inflammatory Activity
    Niosomal formulation of diclofenac sodium prepared with 70% cholesterol
     showed greater anti-inflammatory effect as compared to the free drug
    Similarly, nimesulide and flurbiprofen showed greater activity than the free
     drug
                                                                                52
3.For Targeting and Retention of Drug in
   Blood Circulation
3.1 For Increased Uptake by A431 Cells [a model cell                          line (epidermoid
   carcinoma) used in biomedical research]
    Chitosan based vesicles incorporating transferrin and glucose as ligand have been reported.
     These vesicles bind CoA (co-A) to their surface. Chitosan containing vesicles are then taken
     up by A431 cells and the uptake was found to be enhanced by transferrin.
3.2 For Liver Targeting
    Methotrexate was reported to be selectively taken up by liver cells after administration as a
     niosomal drug delivery system.
3.3 To Improve the Efficacy of Drugs in Cancer Therapy
    Niosomes can also be used as a suitable delivery system for the administration of drugs like
     5-FU.
    Niosomes of doxorubicin prepared from C16 monoalkyl glycerol ether with or without
     cholesterol, exhibited an increased level of doxorubicin in tumor cells, serum and lungs, but
     not in liver and spleen
    Niosomal preparation of methotrexate exhibited greater antitumor activity as compared to
     plain drug solution
                                                                                             53
3.4 In Treatment of Localized Psoriasis
     In the treatment of localized psoriasis, niosomes of methotrexate taking
      chitosan as polymer have shown promising results
3.5 In Leishmaniasis
    The leishmaniasis parasite mainly infects liver and spleen cells. The commonly used
     drugs, antimonials, may damage the body organ like heart, liver, kidney etc. The
     efficiency of sodium stibogluconate has been found to be enhanced by incorporation in
     niosomes. The additive effect was observed for two doses given on successive days.
     Moreover, the distribution of antimony in mice showed the higher level of antimony in
     liver after its intravenous (i.v.) administration via niosomes drug formulation.
3.6 In Diagnostic Imaging
    It has been studied that niosomes can also act as a carrier radiopharmaceuticals and
     showed site specificity for spleen and liver for their imaging studies using 99mTc labelled
     DTPA (diethylene triamine pentaacetic acid) containing niosomes.
    Conjugated niosomal formulations of gadobenate with (N-palmitoyl-glucosamine,
     NPG), PEG 4400 and both PEG and NPG can be used to increased tumor targeting of a
     paramagnetic agent

                                                                                           54
3.7 Carrier for Haemoglobin
     Niosomes play an important role as a carrier for haemoglobin. The
      niosomal haemoglobin suspension was found to give superimposable
      curve on free haemoglobin curve
                    Usefulness of Niosomes in Cosmetics
 Niosomes of N-acetyl glucosamine are prepared due to its potential in the delivery of
  hydrophilic and hydrophobic drugs in topical form and improved penetration into the
  skin. Prepared formulations improved the extent of drug localized in the skin, as
  needed in hyperpigmentation disorders.
 Elastic niosomes showed increased permeation through the skin which will be
  beneficial for topical anti-aging application.
 suitable for skin moisturising and tanning products
 Niosomes were prepared as possible approach to improve the low skin penetration and
  bioavailability shown by conventional topical vehicle for minoxidil.
 Niosomes with added solubilizers enhanced the permeation of ellagic acid (a potent
  antioxidant phytochemical substance which has limited use due to poor
  biopharmaceutical properties, low solubility and low permeability) into the skin with
                                                                                   55
  increased efficacy of ellagic acid
56
57
58
Marketed products
Lancome has come out with a variety of anti-ageing
 products which are based on niosome formulations.
 L’Oreal is also conducting research on anti-ageing
 cosmetic products. A picture of the Lancome anti-ageing
 formulation is below




                       LANCOME FOUNDATION CREAM NIOSOME+
                                                           59
Future Prospects
• Niosomes      represent      a    promising      drug      delivery
  module.
• There is lot of scope to encapsulate toxic anti-cancer drugs, anti
  infective drugs, anti AIDS drugs, anti-inflammatory drugs, anti
  viral drugs, etc. in niosomes and to use them as promising drug
  carriers to achieve better bioavailability and targeting properties
  and for reducing the toxicity and side effects of the drugs.
• The ionic drugs carriers are relatively toxic and        unsuitable
  whereas niosomal carriers are safer.
                                                                  60
• In addition, handling and storage of niosomes require no special

  conditions.

• Vesicular drug carriers like niosomes can be transported by

  microphages which are known to infiltrate tumour cells.

• It may be possible to take advantage of these activated macrophage

  system in delivering the anti tumour agents within vesicles more

  quantitatively to tumour sites.
                                                               61
Conclusion
In nutshell, as a drug delivery device, compared to liposomes,
 niosomes are osmotically active and are quite stable chemically by
 their own as well as improve the stability of the drug so entrapped
 and delivered. They do not require special conditions for handling,
 protection or storage and industrial manufacturing. Beside this,
 they offer flexibility in structural characteristics (composition,
 fluidity, size), and can be designed as desired.
Niosomes offer various advantages over other drug delivery
 devices and have found applicability in pharmaceutical field.
It was thus concluded that niosomes are very effective drug
 delivery tools for incorporation/targeting of various therapeutically
 active moieties and the onus lies on future scientists to effectively
 harness its potential in diverse application areas for the benefit of
 mankind.                                                          62
References
1. Sanjay K. Jain and N.K. Jain Controlled and novel drug delivery
system

2. Dr. Rakesh S. Patel niosomes as a unique drug delivery system,
www.pharmainfo.net

3. Mithal, B. M., A text book of pharmaceutical formulation, 6 th
Edn., vallabh prakashan, 6, 306-307

4. International journal of pharmaceutical Science            and
Nanotechnology volume 1,issue 1, April-June 2008

5. Shailendra Kumar Singh et.al, Niosomes: A Controlled and
Novel Drug Delivery System, Biol. Pharm. Bull. 34(7) 945—953
(July, 2011)                                                 63

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Niosome

  • 1. By Dr. Shreeraj Shah Associate Professor, Dept. of Pharmaceutical Technology, L.J. Institute of Pharmacy, Ahmedabad 1
  • 2. Content Introduction General characteristics of Niosomes Advantage of Niosomes Disadvantage of Niosomes Structure of Niosomes Contrast and similarity between Niosomes and Liposomes Methods of Preparation Factors affecting the physicochemical properties of Niosomes Types of Niosomes Characterization of Niosomes Therapeutic Applications Marketed products Future Prospects Conclusion 2 References
  • 3. Introduction • Development of new drug, improving safety and efficacy of existing drugs is difficult, expensive and time consuming • At present, no available drug delivery system behaves ideally achieving all the lofty goals • Encapsulation of the drug in vesicular structures is one of the promising system • such as liposomes, niosomes, transferosomes, ethosomes and pharmacosomes etc • It delivers drug directly to the site of action, leading to reduction of drug toxicity with no adverse effects 3
  • 4. •Vesicular drug delivery reduces the cost of therapy by improving bioavailability of medication and also solves the problems of drug insolubility, instability and rapid degradation •Niosomes are a novel drug delivery system, in which the medication is encapsulated in a vesicle, composed of a bilayer of non-ionic surface active agents and hence the name niosomes •The niosomes are very small, and microscopic in size. Their size lies in the nanometric scale 4
  • 5. Definition • Niosomes are synthetic microscopic vesicles consisting of an aqueous core enclosed in a bi layer consisting of cholesterol and one or more nonionic surfactants • Vesicles are prepared from self assembly of hydrated non ionic surfactants molecules 5
  • 6. General Characteristics of Niosome •Biocompatible, biodegradable, non-toxic, non immunogenic and non-carcinogenic •The ability of nonionic surfactant to form bilayer vesicles is dependant on the HLB value of the surfactant, the chemical structure of the components and the critical packing parameter •Niosomes can be characterized by their size distribution studies • High resistance to hydrolytic degradation •The properties of niosome depends both on composition of the bilayer & on method of their production 6
  • 7. Advantages of Niosomes  Targeted drug delivery can be achieved using niosomes the drug is delivered directly to the body part where the therapeutic effect is required  Reduced dose is required to achieve the desired effect  Subsequent decrease in the side effects  The therapeutic efficacy of the drugs is improved by reducing the clearance rate, targeting to the specific site and by protecting the encapsulated drug  Niosomes are amphiphillic i.e. both hydrophilic and lipophillic in nature and can accommodate a large number of drugs with a wide range of solubilities  Improve the oral bioavailability of poorly soluble drugs  Enhance the skin permeability of drugs when applied topically  provide advantage of usage through various routes viz. oral, parentral, topical, ocular etc.  The bilayers of the niosomes protect the enclosed Active pharmaceutical ingredient from the various factors present both inside and outside the body  The surfactants used and also the prepared niosomes are biodegradable, biocompatible and non-immunogenic  They are osmotically active and stable 7
  • 8. Disadvantages of Niosome •Aqueous suspension of niosome may exhibit fusion, aggregation leaching or hydrolysis of entrapped drug, thus limiting the shelf life of niosome dispersion. • Time consuming • Requires specialized equipment • Inefficient drug loading 8
  • 9. Structure of Niosomes •Niosomes are microscopic lamellar structures •Basic structural components are  Non ionic surfactant  Cholesterol  Charge inducing molecule •A number of non-ionic surfactants used are: polyglycerol alkyl ether, glucosyl dialkyl ethers, crown ethers, ester linked surfactants, polyoxyethylene alkyl ether and a series of spans and tweens 9
  • 10. Non-ionic Surfactants The non-ionic surfactants orient themselves in bilayer lattices where the polar or hydrophobic heads align facing aqueous bulk (media) while the hydrophobic head or hydrocarbon segments align in such a way that the interaction with the aqueous media would be minimized.  To attain thermodynamic stability, every bilayer folds over itself as continuous membrane i.e. forms vesicles so that hydrocarbon/water interface remains no more exposed. 10
  • 11. Mainly following types of non-ionic surfactants are used for the formation of niosomes:  Alkyl Ethers: L’Oreal described some surfactants for the preparation of niosomes containing drugs/chemicals as  1) Surfactant-I (molecular weight (MW 473)) is C16 monoalkyl glycerol ether with average of three glycerol units.  2) Surfactant-II (MW 972) is diglycerol ether with average of the seven glycerol units.  3) Surfactant III (MW 393) is ester linked surfactant. Other than alkyl glycerol, alkyl glycosides and alkyl ethers bearing polyhydroxyl head groups are also used in formulation of niosomes.  Alkyl Esters: Sorbitan esters are most preferred surfactant used for the preparation of niosomes amongst this category of surfactants. Vesicles prepared by the polyoxyethylene sorbitan monolaurate are relatively soluble than other surfactant vesicles. For example polyoxyethylene (polysorbate 60) has been utilized for encapsulation of diclofenac sodium. A mixture of polyoxyethylene-10-stearyl ether : glyceryl laurate : cholesterol (27 : 15 : 57) has been used in transdermal delivery of cyclosporine-A.  Alkyl Amides: Alkyl amide (e.g. galactosides and glucosides) have been utilized to produce niosomal vesicles.  Fatty Acid and Amino Acid Compounds: Long chain fatty acids and amino acid moieties have also been used in some niosomes preparation. 11
  • 12. Diff. types of Non-ionic surfactants 12
  • 13. Cholesterol Steroids are important components of the cell membrane and their presence in membrane affect the bilayer fluidity and permeability. Cholesterol is a steroid derivative, which is mainly used for the formulation of niosomes. Although it may not show any role in the formation of bilayer, its importance in formation of niosomes and manipulation of layer characteristics can not be discarded. In general, incorporation of cholesterol affects properties of niosomes like membrane permeability, rigidity, encapsulation efficiency, ease of rehydration of freeze dried niosomes and their toxicity. It prevents the vesicle aggregation by the inclusion of molecules that stabilize the system against the formation of aggregates by repulsive steric or electrostatic forces that leads to the transition from the gel to the liquid phase in niosome systems. As a result of this, the niosome becomes less leaky in nature. 13
  • 14. Charge inducing molecule Some charged molecules are added to niosomes to increase stability of niosomes by electrostatic repulsion which prevents aggregation and coalescence. The negatively charged molecules used are diacetyl phosphate (DCP) and phosphotidic acid. Similarly, stearylamine (STR) and stearyl pyridinium chloride are the well known positively charged molecules used in niosomal preparations. Only 2.5-5 mol % concentration of charged molecules is tolerable because high concentration can inhibit the niosome formation 14
  • 15. 15
  • 16. Contrast of Niosomes Vs liposomes Niosome Liposome 1. Less Expensive 1. More expensive 2. Chemically Stable 2. Chemically unstable 3. Niosomes are prepared 3. liposomes are prepared from uncharged single- from double-chain chain surfactant phospholipids 4. They do not require 4. They require special special storage and storage, handling & handling purity of natural phospholipid is variable. 5. Non ionic drugs carriers 5. The ionic drugs carriers are safer are relatively toxic & unsuitable 16
  • 17. Similarity of niosomes and liposomes • However Niosomes are similar to liposomes in functionality. • Niosomes also increase the bioavailability of the drug and reduce the clearance like liposomes. • Niosomes can also be used for targeted drug delivery, similar to liposomes. • As with liposomes, the properties of the niosomes depend both- on the composition of the bilayer, and the method of production used. 17
  • 18. Method of Preparation 1. Ether Injection (LUV) Based on the vesicle size, niosomes can be divided into three groups. 2. Hand Shaking Method (MLV) These are small unilamellar vesicles (SUV, size=0.025-0.05 μm), 3. The “Bubble” Method multilamellar vesicles (MLV, size=>0.05 μm), and large 4. Reverse Phase Evaporation (LUV) unilamellar vesicles (LUV, size=>0.10 μm). 5. Sonication (SUV) 6. Multiple membrane extrusion method 7. Trans Membrane pH Gradient Drug Uptake Process (remote Loading) (MLV) 8. Microfluidization method (SUV) 9. Formation of Niosomes From Proniosomes 18
  • 19. General steps of Niosome preparation • Hydration of mixture of surfactant/lipid at elevated temperature • Sizing of niosomes • Removal of unentrapped material from vesicles 19
  • 20. Common stages of all methods of preparation of Niosomes Cholesterol + Non ionic surfactant Dissolve in organic solvent Solution in organic solvent Drying Thin film Dispersion (Hydration) Niosome suspension 20
  • 21. 1. Ether injection method • This method is based on slow injection of surfactant : cholesterol solution in ether through 14 gauge needle into a preheated aqueous phase maintained at 600C • Vaporization of ether resulting into a formation of ether gradient at ether-water interface which leads to formation of single layered vesicles • Depending upon the conditions used, the diameter of the vesicle range from 50 to 1000 nm 21
  • 22. 22
  • 23. 2. Hand shaking method (Thin film hydration technique) • Surfactant and cholesterol are dissolved in a volatile organic solvent (diethyl ether, chloroform or methanol) in a round bottom flask • The organic solvent is removed under vaccum at room temperature using rotary evaporator leaving a thin layer of solid mixture deposited on the wall of the flask • The dried surfactant film can be rehydrated with aqueous phase at temperature slightly above the phase transition temperature of the surfactant used, with gentle agitation • This process forms large multilamellar niosomes 23
  • 24. 24
  • 25. 3. The “Bubble” Method •It is novel technique for the one step preparation of liposomes and niosomes without the use of organic solvents •The bubbling unit consists of round-bottom flask with three necks positioned in water bath to control the temperature •Water-cooled reflux and thermometer is positioned in the first and second neck and nitrogen supply through the third neck • Cholesterol and surfactant are dispersed together in this buffer (pH 7.4) at 70°C, the dispersion mixed for 15 seconds with high shear homogenizer and immediately afterwards “bubbled at 70°C using nitrogen gas 25
  • 26. 4. Reverse Phase Evaporation Technique (REV) •Cholesterol and surfactant (1:1) are dissolved in a mixture of ether and chloroform •An aqueous phase containing drug is added to this and the resulting two phases are sonicated at 4-5°C. The clear gel formed is further sonicated after the addition of a small amount of phosphate buffered saline (PBS) •The organic phase is removed at 40°C under low pressure. The resulting viscous niosome suspension is diluted with PBS and heated on a water bath at 60°C for 10 min to yield niosomes 26
  • 27. 27
  • 28. 5. Sonication •In this method, an aliquot of drug solution in buffer is added to the surfactant/cholesterol mixture in a 10-ml glass vial. •The mixture is probe sonicated at 60°C for 3 minutes using a sonicator. •The resultant vesicles are of small unilamellar type niosomes. 6. Multiple membrane extrusion method • In this method, a mixture of surfactant, cholesterol and diacetyl phosphate is prepared and then solvent is evaporated using rotary vacuum evaporator to leave a thin film. The film is then hydrated with aqueous drug solution and the suspension thus obtained is extruded through the polycarbonate membrane (mean pore size 0.1 mm) and then placed in series up to eight passages to obtain uniform size niosomes. •Good method of controlling niosome size. 28
  • 29. Sonication method Extrusion method 29
  • 30. 7. Trans membrane pH gradient (inside acidic) Drug Uptake Process (remote Loading) •Surfactant and cholesterol are dissolved in chloroform • The solvent is then evaporated under reduced pressure to get a thin film on the wall of the round bottom flask •The film is hydrated with 300 mM citric acid (pH 4.0) by vortex mixing. •The multilamellar vesicles are frozen and thawed 3 times and later sonicated. To this niosomal suspension, aqueous solution containing 10 mg/ml of drug is added and vortexed • The pH of the sample is then raised to 7.0-7.2 with 1M disodium phosphate • This mixture is later heated at 60°C for 10 minutes to give niosomes 30
  • 31. 8. Microfluidization Method In this method two fluidized streams (one containing drug and the other surfactant) interact at ultra high velocity, in precisely defined micro channels within the interaction chamber in such a way that the energy supplied to the system remains in the area of niosomes formations. This is called submerged jet principle. It results in better uniformity, smaller size and reproducibility in the formulation of niosomes 31
  • 32. 9. Formation of Niosomes from Proniosomes •Another method of producing niosomes is to coat a water-soluble carrier such as sorbitol with surfactant • The result of the coating process is a dry formulation. In which each water-soluble particle is covered with a thin film of dry surfactant. This preparation is termed “Proniosomes” •The niosomes are formed by the addition of aqueous phase at T > T m and brief agitation T = Temperature. Tm = mean phase transition temperature 32
  • 33. Separation of unentrapped drugs The removal of unentrapped solute from the vesicles can be accomplished by various techniques, which include: Gel Filtration The unentrapped drug is removed by gel filtration of niosomal dispersion through a Sephadex-G-50 column and elution with phosphate buffered saline or normal saline. Dialysis The aqueous niosomal dispersion is dialyzed in a dialysis tubing against phosphate buffer or normal saline or glucose solution Centrifugation The niosomal suspension is centrifuged and the supernatant is separated. The pellet is washed and then resuspended to obtain a niosomal suspension free from unentrapped 33 drug.
  • 34. Factors affecting the physicochemical properties of niosomes Membrane Additives Stability of niosomes can be increased by the number of additives into niosomal formulation along with surfactant and drugs. The membrane stability, morphology and permeability of vesicles are affected by numbers of additives e.g. addition of cholesterol in niosomal system increases the rigidity and decreases the drugs permeability through the membrane Temperature of Hydration Shape and size of niosome is also influenced by the hydration temperature. Assembly of the niosomes vesicles is affected by the temperature change of niosomal system. Temperature change can also induce the vesicle shape transformation 34
  • 35. Properties of Drugs  The drug entrapment in niosomes is affected by molecular weight, chemical structure, hydrophilicity, lipophilicity as well as the hydrophilic lipophilic balance (HLB) value of the drug. Vesicle size may increase due to entrapment of drug. Drug particle interact with the surfactant head groups, which may increase charge on polymer and thus cause repulsion of the surfactant bilayer which leads to increase in vesicle size. Amount and Type of Surfactant  As the HLB value of surfactants like span 85 (HLB 1.8) to span 20 (HLB 8.6) increased, the mean size of niosomes also increases proportionally. It is due to the fact that surface free energy decreases with increase in hydrophilicity of surfactant. Alkyl chain is present in well ordered structure in gel state, while in the liquid state the structure of bilayer is more disordered.  Entrapment efficiency is also affected by phase transition temperature i.e. span 60 having higher TC, provide better entrapment efficiency. Entrapment efficiency of the niosomes is affected by the HLB value for e.g. niosomes have high entrapment efficiency at HLB value 8.6 but HLB value 14 to 17 is 35 not suitable for niosomes formulation
  • 36. Effect of the nature of drug on the formation of niosomes 36
  • 37. Cholesterol Content and charge  Hydrodynamic diameter and entrapment efficiency were found to be increased due to cholesterol content in the niosomal bilayer. Cholesterol can act by two ways. First, it can increase the chain order of liquidated bilayer and second, by decreasing the chain order of the gel state bilayer.  Release rate of drug decreases and rigidity of bilayer increases due to high concentration of cholesterol  The presence of charge tends to increase the interlamellar distance between successive bilayers in multilamellar vesicle structure and leads to greater overall entrapped volume Structure of Surfactants  The geometry of vesicles to be formed from surfactant is affected by its structure, which is related to critical packing parameter (CPP) where V= hydrophobic group volume Ic = the critical hydrophobic group length a0 = the area of hydrophilic head group spherical micelles formed if CPP<0.5, inverted micelles is formed if CPP>1. 37
  • 38. Method of Preparation  The average size of acyclovir niosomes prepared by hand-shaking process was larger (2.7m m) as compared to the average size of niosomes 1.5m m prepared by ether injection method which may be attributed to the passage of cholesterol and span-80 solution through an orifice into the drug solution. Reverse phase evaporation can be used to produce smaller size vesicles. Vesicles with smaller size and greater stability can be produced by microfluidization method. Niosomes obtained by transmembrane pH gradient (inside acidic) drug uptake process showed greater entrapment efficiency and better retention of drug Resistance to Osmotic Stress  Diameter of niosomal vesicles was found to be decreased when niosomal suspension is kept in contact with hypertonic salt solution. There is slow release with slight swelling of vesicles, which is due to inhibition eluting fluids from vesicles, followed by faster release, which may be due to decrease in mechanical strength under osmotic stress 38
  • 39. Types of Niosomes Proniosomes Proniosomes are the niosomal formulation containing carrier and surfactant, which requires to be hydrated before being used. The hydration results in the formation of aqueous niosome dispersion. Proniosomes decreases the aggregation, leaking and fusion problem associated with niosomal formulation Aspasomes Combination of acorbyl palmitate, cholesterol and highly charged lipid diacetyl phosphate leads to the formation of vesicles called aspasomes. Aspasomes are first hydrated with water/aqueous solution and then sonicated to obtain the niosomes. Aspasomes can be used to increase the transdermal permeation of drugs. Aspasomes have also been used to decrease disorder caused by reactive oxygen species as it has inherent antioxidant property 39
  • 40. Vesicles in Water and Oil System (v/w/o) It has been reported that the emulsification of an aqueous niosomes into an oil phase form vesicle in water in oil emulsion (v/w/o). This can be prepared by addition of niosomes suspension formulated from mixture of sorbitol monostearate, cholesterol and solulan C24 (poly-24-oxyethylene cholesteryl ether) to oil phase at 60°C. This results in the formation of vesicle in water in oil (v/w/o) emulsion which by cooling to room temperature forms vesicle in water in oil gel (v/w/o gel). The v/w/o gel thus obtained can entrap proteins/proteinous drugs and also protect it from enzymatic degradation after oral administration and controlled release. The immunogenic properties of v/w/o gel and w/o gel reported that both exhibit immunoadjuvant tendency. In this system aqueous niosomes (v/w) are emulsified into the oily phase 40
  • 41. Niosomes in Carbopol Gel Niosomes were prepared using drug, spans and cholesterol. The niosomes thus obtained were then incorporated in carbopol-934 gel (1% w/w) base containing propylene glycol (10% w/w) and glycerol (30% w/w). It was observed that the mean flux value and diffusion co-efficient were 5 to 7 times lower for niosomal gel as compared to plain drug gels. Moreover, carrageenan induced paw edema inhibition (i.e. 66.68 ± 5.19%) was higher by niosome formulation as compared to plain gel. Niosomes of Hydroxyl Propyl Methyl Cellulose In this type, a base containing 10% glycerin of hydroxy propyl methyl cellulose was first prepared and then niosomes were incorporated in it. The bioavailability and reduction of paw edema induced by carrageenan was found to be higher by this niosomal system than the plain formulation of flurbiprofen. 41
  • 42. Characterization of niosomes Physical characterization Chemical characterization Biological characterization 42
  • 43. Physical characterization Sr no. Parameters Analytical methods/Instruments 1. Vesicles shape & surface Transmission electron morphology microscopy, Freeze-fracture electron microscopy 2. Mean vesicle size and size Dynamic light scattering, distribution zetasizer, (submicron and micron range) Photon correlation spectroscopy, laser light scattering, gel permeation and gel exclusion 3. Surface charge Free-flow electrophoresis 43
  • 44. 4. Phase behavior Freeze-fracture electron microscopy, Differential scanning colorimetery 5. Electrical surface potential Zeta potential and surface pH measurements & pH sensitive probes 6. Entrapment efficiency Amount entrapped/total amount added*100 44
  • 45.  Chemical characterization Sr no Parameters Analytical methods/Instruments 1. Cholesterol concentration Cholesterol oxidase assay and HPLC 2. Cholesterol auto-oxidation HPLC and TLC 3. Osmolarity Osmometer 45
  • 46.  Biological characterization Sr No. Parameters Analytical methods/Instruments 1. Sterility Aerobic or anaerobic cultures 2. Pyrogenicity Limulus Amebocyte Lysate (LAL) test 3. Animal toxicity Monitoring survival rates, histology and pathology 46
  • 47. Evaluation Parameters (Summary) % Entrapment Efficiency (%EE) Size, Shape and Morphology  Transmission Electron Microscopy (TEM)  Freeze Fractured Microscopy  Optical Microscopy Technique In Vitro Release Study Tissue Distribution/In Vivo Study Stability Study Number of Lamellae Membrane Rigidity Vesicular Surface Charge 47
  • 48. Drugs and routes of administration used in Niosomes 1. Intravenous Route Ipromide, Vincristine,Indomethacin, Colchicines, Rifampicin, Transferrin, Zidovudine, Cisplantin, amarogentin, Daunorubicin, AmphotericinB. 2. Transdermal Route Flurbiprofen, Piroxicam, Levonorgestrol, Nimeluside, Estradiol, Ketoconazole, Enoxacin, DNA loaded noisome, Cyclosporine, Erythromycin, α interferon. 3. Oral Route Vaccine, Polysaccharide Coated noisome, Cipro floxacin, Insulin. 4. Oncology Route Methotrexate, Doxorubicin, adriamycin. 5. Ocular Route Timolol, cyclopentolate 6. Nasal Route Sumatriptan, Influenza 7.Immunological Adjuvant Bovine serum albumin, Hemoglobin 8. For treatment of Leishmaniasis Stilbogluconat 48
  • 49. Therapeutic applications of Niosomes The applications of niosomes can be mainly classified into three categories 1) For Controlled Release of Drugs 2) To Improve the Stability and Physical Properties of the Drugs 3) For Targeting and Retention of Drug in Blood Circulation 49
  • 50. 1)To Prolong the Release Rate of Drugs 1.1 For Controlled Release The release rate of drugs like withaferin and gliclazide from the niosomes was found slower as compared to other dosage forms 1.2 In Ophthalmic Drug Delivery Experimental results of the water soluble antibiotic gentamicin sulphate showed a substantial change in the release rate. Beside this, the percent entrapment efficiency of gentamicin sulphate was altered when administered as niosomes. Also, as compared to normal drug solution, niosomes of drug show slow release Niosomal formulation containing timolol maleate (0.25%) prepared by chitosan coating exhibited more effect on intra ocular tension with fewer side effects as compared to the marketed formulation. 50
  • 51. 2) To Improve the Stability and Physical Properties of the Drugs 2.1 To Increase Oral Bioavailability  With the formulation of niosomes, the oral bioavailability of the acyclovir as well as griseofulvin was increased as compared to the drug alone. Similarly, the absorptivity of poorly absorbed peptide and ergot alkaloid can be increased by the administration in the bile duct of rats when administered as micellar solution together with the POE-24-cholesteryl ester 2.2 For Improvement of Stability of Peptide Drugs  8-arginin vasopressin, 9-glycinamide-ω  The in vitro release of insulin from niosomes formulated by span 40 and span 60 in simulated intestinal fluid was lower than the niosomes formulated by span 20 and span 80.  Niosomes prepared by the span 60 has high resistance against proteolytic enzyme and exhibit good stability in storage temperature and in presence of sodium deoxycholate 51
  • 52. 2.3 To Promote Transdermal Delivery of Drugs  Many drugs such as lidocaine, estradiol, cyclosporine etc. are used for topical and transdermal drug delivery system by formulating them as niosomes  The niosomes of natural compound, ammonium glycyrrhizinate were formulated for effective anti-inflammatory activity using new non-ionic surfactant, bola surfactat-span 80-cholesterol (2 : 3 : 1 ratio).  Experimental study showed that the bola niosomes were able to promote the intracellular uptake of ammonium glycerrhizinic acid 2.4 As a Tool for Improvement of Stability of Immunological Products  Important tool for immunological selectivity, low toxicity and more stability of the incorporated active moiety 2.5 To Improve Anti-inflammatory Activity  Niosomal formulation of diclofenac sodium prepared with 70% cholesterol showed greater anti-inflammatory effect as compared to the free drug  Similarly, nimesulide and flurbiprofen showed greater activity than the free drug 52
  • 53. 3.For Targeting and Retention of Drug in Blood Circulation 3.1 For Increased Uptake by A431 Cells [a model cell line (epidermoid carcinoma) used in biomedical research]  Chitosan based vesicles incorporating transferrin and glucose as ligand have been reported. These vesicles bind CoA (co-A) to their surface. Chitosan containing vesicles are then taken up by A431 cells and the uptake was found to be enhanced by transferrin. 3.2 For Liver Targeting  Methotrexate was reported to be selectively taken up by liver cells after administration as a niosomal drug delivery system. 3.3 To Improve the Efficacy of Drugs in Cancer Therapy  Niosomes can also be used as a suitable delivery system for the administration of drugs like 5-FU.  Niosomes of doxorubicin prepared from C16 monoalkyl glycerol ether with or without cholesterol, exhibited an increased level of doxorubicin in tumor cells, serum and lungs, but not in liver and spleen  Niosomal preparation of methotrexate exhibited greater antitumor activity as compared to plain drug solution 53
  • 54. 3.4 In Treatment of Localized Psoriasis  In the treatment of localized psoriasis, niosomes of methotrexate taking chitosan as polymer have shown promising results 3.5 In Leishmaniasis  The leishmaniasis parasite mainly infects liver and spleen cells. The commonly used drugs, antimonials, may damage the body organ like heart, liver, kidney etc. The efficiency of sodium stibogluconate has been found to be enhanced by incorporation in niosomes. The additive effect was observed for two doses given on successive days. Moreover, the distribution of antimony in mice showed the higher level of antimony in liver after its intravenous (i.v.) administration via niosomes drug formulation. 3.6 In Diagnostic Imaging  It has been studied that niosomes can also act as a carrier radiopharmaceuticals and showed site specificity for spleen and liver for their imaging studies using 99mTc labelled DTPA (diethylene triamine pentaacetic acid) containing niosomes.  Conjugated niosomal formulations of gadobenate with (N-palmitoyl-glucosamine, NPG), PEG 4400 and both PEG and NPG can be used to increased tumor targeting of a paramagnetic agent 54
  • 55. 3.7 Carrier for Haemoglobin  Niosomes play an important role as a carrier for haemoglobin. The niosomal haemoglobin suspension was found to give superimposable curve on free haemoglobin curve Usefulness of Niosomes in Cosmetics  Niosomes of N-acetyl glucosamine are prepared due to its potential in the delivery of hydrophilic and hydrophobic drugs in topical form and improved penetration into the skin. Prepared formulations improved the extent of drug localized in the skin, as needed in hyperpigmentation disorders.  Elastic niosomes showed increased permeation through the skin which will be beneficial for topical anti-aging application.  suitable for skin moisturising and tanning products  Niosomes were prepared as possible approach to improve the low skin penetration and bioavailability shown by conventional topical vehicle for minoxidil.  Niosomes with added solubilizers enhanced the permeation of ellagic acid (a potent antioxidant phytochemical substance which has limited use due to poor biopharmaceutical properties, low solubility and low permeability) into the skin with 55 increased efficacy of ellagic acid
  • 56. 56
  • 57. 57
  • 58. 58
  • 59. Marketed products Lancome has come out with a variety of anti-ageing products which are based on niosome formulations. L’Oreal is also conducting research on anti-ageing cosmetic products. A picture of the Lancome anti-ageing formulation is below LANCOME FOUNDATION CREAM NIOSOME+ 59
  • 60. Future Prospects • Niosomes represent a promising drug delivery module. • There is lot of scope to encapsulate toxic anti-cancer drugs, anti infective drugs, anti AIDS drugs, anti-inflammatory drugs, anti viral drugs, etc. in niosomes and to use them as promising drug carriers to achieve better bioavailability and targeting properties and for reducing the toxicity and side effects of the drugs. • The ionic drugs carriers are relatively toxic and unsuitable whereas niosomal carriers are safer. 60
  • 61. • In addition, handling and storage of niosomes require no special conditions. • Vesicular drug carriers like niosomes can be transported by microphages which are known to infiltrate tumour cells. • It may be possible to take advantage of these activated macrophage system in delivering the anti tumour agents within vesicles more quantitatively to tumour sites. 61
  • 62. Conclusion In nutshell, as a drug delivery device, compared to liposomes, niosomes are osmotically active and are quite stable chemically by their own as well as improve the stability of the drug so entrapped and delivered. They do not require special conditions for handling, protection or storage and industrial manufacturing. Beside this, they offer flexibility in structural characteristics (composition, fluidity, size), and can be designed as desired. Niosomes offer various advantages over other drug delivery devices and have found applicability in pharmaceutical field. It was thus concluded that niosomes are very effective drug delivery tools for incorporation/targeting of various therapeutically active moieties and the onus lies on future scientists to effectively harness its potential in diverse application areas for the benefit of mankind. 62
  • 63. References 1. Sanjay K. Jain and N.K. Jain Controlled and novel drug delivery system 2. Dr. Rakesh S. Patel niosomes as a unique drug delivery system, www.pharmainfo.net 3. Mithal, B. M., A text book of pharmaceutical formulation, 6 th Edn., vallabh prakashan, 6, 306-307 4. International journal of pharmaceutical Science and Nanotechnology volume 1,issue 1, April-June 2008 5. Shailendra Kumar Singh et.al, Niosomes: A Controlled and Novel Drug Delivery System, Biol. Pharm. Bull. 34(7) 945—953 (July, 2011) 63