Liposomes

Unit I
M.pharmacy I
Neha Srivastava
 Introduction
Slide Title

• Liposomes are cocentric bilayerd vesicles in

which aquoues volume is entirely enclosed
by a membraneous lipid bilayer.
• 1960 by Bangham et.al.
• Targeting and delivery system
• Natural and synthetic phospholipids
• diameter ranges from 80nm to 100um
 Introduction
Slide Title

Advantages
• Provides selective passive targeting
• Increase efficacy and therapeutic index
• Increase stability via encapsulation
• Reduction in toxicity
• Site avoidance effect
• Improved pharmacokinetics
• Flexibility with site specific legands
 General structure of liposome
.

Lipid bilayer
Mechanism of liposome formation
Slide Title

Physiochemical features of phospholipids

• Ampiphillic molecules (Polar head and non
polar tail)
• Tail 2 Fatty acid chain 10-24 C
and 0-6 double bonds
• Polar end bind to phosphoric acid
• Supramolecular structure
• In water, they spontaneously form closed
structures
Mechanism of liposome formation
Slide Title

Physiochemical features of phospholipids

• Self assembling of structure in aq medium
• Amphiphilic molecules such as soap, detergents
can also be used
• In water they form different microscopic
structures such as lamellar, hexagonal etc
• Commonly used is natural phospholipid-
phosphatidylcholine (PC)
.

Structure of
the
phospholipid
molecule
Mechanism of liposome formation
Slide Title
Phosphatidylcholine

Glycerol group attached polar head and non polar tail

Molecule of PC are not soluble but organize in aq environment
Large free energy shift in aquoeus and hydrophobic environment
State of lowest energy indicates highest stability
Mechanism of liposome formation
Slide Title

Phosphitidylcholine
• Soaps forms spherical micellar structure
• Effect of phase transition temperature
• Hydration of lipid bilayer
• On agitation forms MLV
• Sonication -small unilamilar vesicles
• Mechanical energy – Large unilamilar vesicles
 2. SPHINGOLIPIDS

• They contain sphingosine (or) a related base as

their structural backbone.

• They contain 3 characteristic building blocks:-

- A molecule of fatty acid
- A molecule of sphingosine &
- Head groups that can vary from simple alcohol
as choline to very complex carbohydrates.

• The most abundant sphingolipid in higher animals

is sphingomyelin.
3. GLYCOSPHINGOLIPIDS
• These are found mainly in grey matter of brain tissue
of higher animals and are used in minor component.
• These are included in liposomes formulation to
provide a layer of surface charged groups.

4. STEROLS
• Cholesterol & its derivatives are quite often
included as components of liposomal membrane.
5. SYNTHETIC PHOSPHOLIPIDS
• Saturated phospholipids include dipalmitoyl
phosphatidyl choline(DPPC) , dipalmitoyl
phosphatidyl ethanol amine(DPPE) , dipalmitoyl
phosphatidyl serine(DPPS) .

• Unsaturated phospholipids have also been used which

include dioleo-phosphatidyl choline(DOPC) , dioleo-
phosphatidyl glycerol(DOPG)
6. POLYMERIC MATERIALS

• A large variety of polymerizable lipids which can

form vesicles , has been synthesized.

• These include lipids containing conjugated diene ,

methacrylate , and thiol groups as polymerizable
moieties.
Mechanism of liposome formation
Slide Title
• Effect of molecular geometry
Mechanism of liposome formation
Slide Title
Rigidization of structure
• Cholesterol modulate the bilayer membrane
• Fluidity buffer
• Above phase transition temperature it tends
to make the membrane more order
• Do not form bilayer but can trap in between
• Orientation of cholesterol
• Effect of Phage transition temperature
• 1:1 is efficient
Mechanism of liposome formation
Slide Title
Rigidization of structure
• Cholesterol modulate the bilayer membrane
• Fluidity buffer
• Above phase transition temperature it tends
to make the membrane more order
• Do not form bilayer but can trap in between
• Orientation of cholesterol
• Effect of Phage transition temperature
• 1:1 is efficient
Classification of liposomes
Slide Title
Rigidization of structure
• Cholesterol modulate the bilayer membrane
• Fluidity buffer
• Above phase transition temperature it tends
to make the membrane more order
• Do not form bilayer but can trap in between
• Orientation of cholesterol
• Effect of Phage transition temperature
• 1:1 is efficient
Slide Title
Passive loading Techniques
Mechanical dispersion method of passive
loading
• Lipid solution in organic phase
• Removal of organic phase
• Dry layer formation
• Hydration
• Different parameters such as vortexing,
sonication, etc
Slide Title
Passive loading Techniques

Thin film hydration by hand shaking(MLV)

or Non shaking method (ULV)
Slide Title
Passive loading Techniques

Proliposomes (MLV)
Lipid is dried over the support
Lipid + Support Dry film

Proliposomes Hydration
 Slide Title
Passive loading Techniques

Mechanical treatment of MLVS

Sonication (SUVs)
• Reduce to SUVs
• Two types : Bath and probe
• Probe tip – high energy – liposomal degradation
• Contribute titanium particles
• Sonication for 5-10 min
• Lipid dispersion clarifies
 Slide Title
Passive loading Techniques

Mechanical treatment of MLVS

Sonication (SUVs)
• Reduce to SUVs
• Two types : Bath and probe
• Probe tip – high energy – liposomal degradation
• Contribute titanium particles
• Sonication for 5-10 min
• Lipid dispersion clarifies – further Centrifugation
 Slide Title
Passive loading Techniques

Mechanical treatment of MLVS

Sonication (SUVs)
• Reduce to SUVs
• Two types : Bath and probe
• Probe tip – high energy – liposomal degradation
• Contribute titanium particles
• Sonication for 5-10 min
• Lipid dispersion clarifies – further Centrifugation
Slide Title
Passive loading Techniques
Slide Title
Passive loading Techniques

French pressure cell liposome

• Reduce to SUVs
• Two types : Bath and probe
• Probe tip – high energy – liposomal degradation

• Contribute titanium particles

• Sonication for 5-10 min
• Lipid dispersion clarifies – further Centrifugation
Passive
Slide loading Techniques
Title

French pressure cell liposome

• Uni or oligo lamellar liposomes
• More stable
• Lipid phase composition, temperature and
pressure
• Low leakage
• Less heterogenicity
Passive
Slide loading Techniques
Title

Micro Emulsion Liposome

• Microfludizer SUV
Passive
Slide loading Techniques
Title

Vesicle produced by extrusion techniques

• LUV and MLV
Passive
Slide loading Techniques
Title

Dried reconstituted vesicles

Freeze thaw cycle
Passive
Slide loading Techniques
Title

Liposomes from pre formed vesicles

 pH induced
 Calcium induced
 Cochleate method
 Passive
Slide loading Techniques
Title

SOLVENT DISPERSION METHOD

Ethanol Injection
Passive
Slide loading Techniques
Title

SOLVENT DISPERSION METHOD

Ether Injection
Passive
Slide loading Techniques
Title

SOLVENT DISPERSION METHOD

Ether Injection
 Passive
Slide loading Techniques
Title

Rapid solvent exchange vesicles

Organic solution of lipid is added with syringe
to aq phase under vacuum
Passive
Slide loading Techniques
Title

De-emulsification method
Double emulsion vesicles
Reverse phase evaporation vesicles
Passive
Slide loading Techniques
Title

DETERGENT DEPLETION
• Micelles formation
• Dialysis
• Column chromatography
• Detergent adsorption using bio beads
ActiveTitle
Slide loading Techniques

DETERGENT DEPLETION
• Ion transport
• Weak acid or base
• High encapsulation
• Reduced leaking
• Bed side laoding of drug
ActiveTitle
Slide loading Techniques

• Ion transport
• Weak acid or base
• High encapsulation
• Reduced leaking
• Bed side laoding of drug
Removal
Slide Titleof untrapped drugs

• MLVs compared to LUVs and SUVs

• Centrifugation
• Supernatant contains untrapped drugs
 Characterization
Slide Title of liposomes

Vesicle shape and Lamellarity

• Electron microscope technique

Lamellarity
• Freeze facture electron microscopy
• 31 P nuclear magnetic resonance
 Characterization
Slide Title of liposomes

 Vesicle size and distribution

 Light microscopy
 Fluorescent Microscopy
 Electron microscopy
 TEM
 Lazer light scattering
 Zetasizer
Characterization
Slide Title of liposomes

Surface charge
Encapsulation efficiency
Trapped volume
Phase behaviour