LIPOSOMES

MD ASIF IQBAL
M.PHARM
(PHARMACEUTICS)
JAMIA HAMDARD
 WHAT ARE LIPOSOMES?
 Liposomes are concentric bilayered vescicles
in which an aqueous volume is entirely
enclosed by a membranous lipid bilayer mainly
composed of natural or synthetic
phospholipids.
 Liposomes can be composed of naturally-
derived phospholipids with mixed lipid chains
(like egg phosphatidylethanolamine), or of
pure surfactant components like DOPE
(dioleoylphosphatidylethanolamine).
Liposomes, usually but not by definition,
contain a core of aqueous solution; lipid
spheres that contain no aqueous material are
called micelles.
WHAT ARE LIPOSOMES?
 COMPARISON OF LIPOSOMES AND
NIOSOMES
 Niosomes are non-ionic surfactant based
liposomes. They are mostly formed by
cholesterol incorporation as an
excipient. Other excipients can also be
used.
 They are structurally similar to
liposomes in having a bilayer, however,
the materials used to prepare niosomes
makes them more stable and thus niosomes
offer many more advantages over
liposomes.
ADVANTAGES OF LIPOSOMES
 Provides selective passive targeting to
tumor tissues (e.g. liposomal
doxorubicin)
 Increased efficacy and therapeutic index.

 Increased stability via encapsulation of

drug.
 Reduced toxicity of the encapsulated
agent.
 Improved pharmacokinetics of the drug
(reduced elimination, increased
circulation times).
CLASSIFICATION OF LIPOSOMES
 Liposomesmay be classified by
a wide variety of methods.
 Their nomenclature may also
depend upon:
 the method of preparation,
 structural parameters or
 special function assigned to
them.
CLASSIFICATION OF LIPOSOMES
 One method of classification relies upon
the number of bilayers formed and the
diameter of the resultant vescicles.
 On this basis LIPOSOMES may be CLASSIFIED
as:
 SMALL UNILAMELLAR VESCICLES (SUV’s,
single bilayer, 10-100 nm)
 LARGE UNILAMELLAR VESCICLES (LUV’s,
single bilayer, 100nm-1µm)
 MULTILAMELLAR VESCICLES (MLV’s, several
bilayers, 100nm-20µm)
CLASSIFICATION OF LIPOSOMES
 OLIGOLAMELLAR VESCICLES (OLV’s, more than
one but not as many as MLV’s, 0.1-1µm)
 INTERMEDIATE SIZED UNILAMELLAR VESCICLES
(IUV’s, ~100nm).

Some classification based on method of

preparation include:
 MULTIVESCICULAR VESCICLES (MVV’s, 100nm-
20µm)
 DRIED-RECONSTITUTED VESCICLES (DRV’s, uni
or oligolamellar, ‹1µm)
CLASSIFICATION OF LIPOSOMES
 REVERSE PHASE EVAPORATION VESCICLES
(REV’s, unilamellar, ~0.5µm)
 MICRO-EMULSIFICATION LIPOSOMES
(MEL, multilayered, 0.1-0.2µm)
 LARGE UNILAMELLAR VESCICLES
PREPARED BY EXTRUSION (VET,
single/bilayered, 100nm-1µm)
 STABLE PLURILAMELLAR VESCICLES
(SPLV’s, multilayered, 100nm-2µm)
 MECHANISM OF LIPOSOME
FORMATION
 WHAT ARE PHOSPHOLIPIDS AND THEIR PHYSICO CHEMICAL
PROPERTIES?
 Phospholipids are amphipathic molecules as they
have a hydrophobic tail and an hydrophilic or
polar head.
 The hydrophilic and hydrophobic domains/ segments
within the molecular geometry of amphiphilic
lipids orient and self organise in ordered
supramolecular structure when confronted with
solvents.
 In aqueous medium the molecules in self-assembled
structures are oriented in such a way that the
polar portion of the molecule remains in contact
with the polar environment and at the same time
shields the non-polar part.
 MECHANISM OF LIPOSOME
FORMATION
 In aqueous mixtures these molecules are able
to form various phases, some of them are
stable and others remain in the metastable
state.
 At high concentrations of these polar lipids,
liquid crystalline phases are formed that upon
dilution with excess water can be stabilised
into relatively stable colloidal particles.
 The macroscopic structures often include
lamellar, hexagonal or cubic phases dispersed
as colloidal nanoconstructs referred as
LIPOSOMES, or HEXASOMES or CUBOSOMES,
respectively.
MECHANISM OF LIPOSOME
FORMATION
 MECHANISM OF LIPOSOME
FORMATION
 The amphipathic nature of phospholipids and
their analogues renders them the ability to
form closed concentric bilayers in the
presence of water.
 Liposomes are formed when thin lipid films
or lipid cakes (of amphiphilic nature) are
hydrated and stacks of liquid crystalline
bilayers become fluid and swell.
 The hydrated lipid sheets detach during
agitation and self close to form large,
multi cellular vescicles which prevent the
interaction of water with the hydrocarbon
core of the bilayer at the edges.
 METHODS OF LIPOSOME
PREPARATION AND DRUG
LOADING
 Liposomes are manufactured in majority using
procedures in which the water soluble
(hydrophilic) materials are entrapped using
aqueous solution of these materials as
hydrating fluid or by the addition of
drug/drug solution at some stage during the
manufacture.
 The lipid soluble (lipophilic) materials are
solubilized in the organic solution of the
constitutive lipid(s) and then evaporated to a
dry drug containing lipid film followed by its
hydration.
 METHODS OF LIPOSOME
PREPARATION AND DRUG
LOADING
 Methods of liposome preparation and drug
loading are as follows:
 PASSIVE LOADING TECHNIQUES:

 Mechanical dispersion methods of passive

loading.
 Solvent dispersion methods for passive
loading.
 Detergent depletion methods of passive
loading.
 REMOTE LOADING
PASSIVE LOADING TECHNIQUES
Passive loading technique
involves three different
groups of methods working
on different principles:
 MECHNICAL DISPERSION
 SOLVENT DISPERSION
 DETERGENT SOLUBILIZATION
 MECHANICAL DISPERSION
METHODS OF PASSIVE LOADING
 The methods covered under this category
begin with a lipid solution in organic
solvent and end up with lipid dispersion
in water.
 The various components are typically
combined by co-dissolving the lipids in
an organic solvent.
 The organic solvent is then removed and
the solid lipid mixture is hydrated using
an aqueous buffer.
 The lipids spontaneously swell and
hydrate to form liposomes.
 MECHANICAL DISPERSION
METHODS OF PASSIVE LOADING
The post hydration treatments
include:
 Micro-emulsification
 Sonication
 French pressure cell
 Membrane extrusion
 Dried reconstituted vescicles
 Freeze thawed liposomes.
 SOLVENT DISPERSION METHODS
FOR PASSIVE LOADING
 In solvent dispersion method, lipids
are first dissolved in an organic
solution, which is then brought into
contact with an aqueous phase
containing materials to be entrapped
within the liposome.
 The lipids align themselves at the
interface of organic and aqueous
phase forming monolayer of
phospholipids, which forms half of
the bilayer of the liposome.
 SOLVENT DISPERSION METHODS
FOR PASSIVE LOADING
 Methods employing solvent dispersion can
be categorised on the basis of
miscibility of the organic solvent and
the aqueous phase.
 The various methods used under solvent
dispersion are:
 Ethanol injection

 Ether injection

 Double emulsion vescicles

 Reverse phase evaporation vescicles

 Stable plurilamellar vescicles

DETERGENT DEPLETION METHODS
OF PASSIVE LOADING
 In this method the phospholipids are
brought into intimate contact with the
aqueous phase via detergents, which
associate with the phospholipid molecules
and serve to screen the hydrophobic
portions of the molecule from water.
 The structures so formed are known as
MICELLES and can be composed of several
hundreds of component molecules.
 The size and shape depend upon the
chemical nature of the detergent, their
concentration and other lipids involved.
 REMOTE (ACTIVE) LOADING
 Active loading methods has the following
advantages over the passive encapsulation
methods:
 A high encapsulation efficiency and
capacity.
 A reduced leakage of the encapsulated
compounds.
 Avoidance of biological active compounds
during preparation steps in the
dispersion thus reducing safety hazards.
 REMOTE (ACTIVE) LOADING
 The technique in general applies the
concept of improved loading of drugs due to
various transmembrane gradients, such as
electrical, ionic or specific salt
gradients.
 This technique brings about improved
loading into preformed liposomes using pH
gradients and potential difference across
liposomal membranes.
 A concentration difference in proton
concentration across the membrane of
liposomes can drive the loading of
amphipathic molecules.
 CHARACTERIZATION OF
LIPOSOMES
 Characterization of liposomes is done to
ensure their in vitro and in vivo
performances.
 The different parameters that are
characterised are:
 Vescicle shape and lamellarity

 Vescicle size and size distribution

 Surface charge

 Encapsulation efficiency and trapped

volume
 Chemical characterization of liposomes.
 APPLICATIONS OF LIPOSOMES
 Liposomes have been used for the
following therapeutic and pharmaceutical
applications:
 Liposomes as drug/protein delivery
vehicles.
 Controlled and sustained drug release in
situ.
 Altered pharmacokinetics and
biodistribution.
 Enzyme replacement therapy and lysosomal
storage disorders.
 Enhanced drug solubilization.
APPLICATION OF LIPOSOMES
 Liposomes in anti microbial, antifungal
and antiviral therapy.
 Liposomal drugs

 Liposome biological response modifiers

 Liposomes in tumor therapy.

 Carriers of small cytotoxic molecules

 Vehicles for macromolecules as cytokines

or genes.
 Liposomes in gene therapy.

 Gene and antisense therapy

 Genetic vaccination
 APPLICATIONS OF LIPOSOMES
 Liposomes in immunology.
 Immunoadjuvant

 Immonumodulator

 Immunodiagnosis

 Liposomes as artificial blood surrogates

 Liposomes as radiopharmaceuticals and

radio diagnostic carriers.
 Liposomes in cosmetics and dermatology

 Lipocomes in enzyme immobilization and

bioreactor technology.
SOME RECENT APPLICATIONS
 Astaxanthine (AST), a red colored
carotenoid pigment, possesses extremely
powerful antioxidative activity. However,
its drawbacks reside in poor solubility
in aqueous system, resulting in extremely
low bioavailability. To ameliorate such
defects, AST encapsulated within
liposomes (AST-L) were prepared. AST-L
apparently showed improved stability and
transportability. The overall transport
time was 7.55 h and 6.00 h for free AST
and AST-L, respectively.
SOME RECENT APPLICATIONS
 Many antibacterial agents, including the glycopeptides, are inactive
against Gram-negative bacteria because of their inability to cross the
outer membrane of these cells. Different chemical and technological
approaches have been described to circumvent such limitation. In this
study, we aimed to apply the strategy of fusogenic liposomes, up to now
used to carry biological compounds and materials inside cells, to localise
a glycopeptide antibiotic, vancomycin (VAN), to the periplasmic space, thus
allowing it to exert its bactericidal activity. Small unilamellar liposome
vesicles were prepared by an extrusion procedure (SUVETs) from a
phospholipid-cholesterol hemisuccinate mixture known for its fusogenic
properties with the eukaryotic cell membrane. VAN was loaded with high
efficiency into these vesicles and in microbiological experiments in vitro
was shown to be able to inhibit to a different extent the growth of wild
and standard Gram-negative bacterial strains. Minimum inhibitory
concentrations as low as 6mg/L were observed, for instance against clinical
isolates of Escherichia coli and Acinetobacter baumannii. In comparison,
neither the free antibiotic nor VAN-loaded 'classical' (non-fusogenic)
liposomes showed any activity against the same bacteria. Scanning and
transmission electron microscopy studies allowed confirmation that the
produced SUVETs were able to adhere to and fuse with the external membrane
of E. coli. According to preliminary experiments, this technological
strategy can be proposed as a potentially successful way to enlarge the
spectrum of activity of VAN.
 REFERENCES
 Vyas, S. P, Khar, R. K, Targeted and
Controlled Drug delivery.
 www.wikipedia.org

 “Improved membrane transport of

astaxanthine by liposomal encapsulation.”
Peng CH, Chang CH, Peng RY, Chyau CC.
 “Encapsulation in fusogenic liposomes
broadens the spectrum of action of
vancomycin against Gram-negative
bacteria.”
Nicolosi D, Scalia M, Nicolosi VM,
Pignatello R.
Thank you!!