1. 1
Krishna S. Jadhav
Department of Pharmaceutics
NATIONAL INSTITUTE OF PHARMACEUTICAL EDUCATION AND RESEARCH
LIPID POLYMER HYBRID NANOPARTICLES
(LPHNPs)
2. Flow of presentation
Introduction
Characteristic of LPHNPs
Types of LPHNPs
Composition of LPHNPs
Method of preparation
Characterization of LPHNPs
Recent advancements in LPHNPs preparation
Application of LPHNPs
Summary 2
3. POLYMERIC NANOPARTICLES
Core shell structured nanoparticles to encapsulate a wide variety of therapeutic agents
The use of amphiphilic polymers results in formation of nanoparticles with a hydrophobic
core and a hydrophilic shell
Polymeric nanoparticles can be prepared from both natural polymers (e.g., chitosan) and
synthetic biodegradable and biocompatible polymers (e.g., poly-lactic-co-glycolic acid
(PLGA)).
4
Advantages
•High structural integrity
Stability during storage
Long systemic circulation half life
Controlled release capability
easy to prepare and readily
functionalized for active targeted
delivery
Disadvantages
•Less biocompatible as
compared to liposomes
4. LIPOSOMES….
Liposomes are simple microscopic vesicles in which an aqueous volume is entirely enclosed by a membrane
composed of lipid molecule
liposomes have long been perceived as the more ideal drug delivery vehicles because of their superior
biocompatibility as liposomes are basically analogues of biological membranes, which can be prepared from
both natural and synthetic phospholipids
Advantages
•Encapsulation of both
hydrophilic and lipophilic drug
Ease of surface modification
Long systemic circulation half
life
Favourable safety
Disadvantages
Lack of structural integrity
Content leakage
Instability during storage
Dose dumping
6. LIPID POLYMER HYBRID NANOPARTICLES
Polymer–lipid hybrid nanoparticles (LPHNPs) are an emerging nanoparticle drug delivery system
made of polymers and lipids taking advantages of both materials
LPHNPs are solid, submicron particles composed of at least two components: the polymer and the
lipid. Various bioactive molecules such as drugs, genes, proteins, and targeting ligands can be
entrapped, adsorbed, or covalently attached in the hybrid system
6
7. Characteristic of LPHNPs
High structural integrity, stability during storage, and controlled release capability attributed to the
polymer core
High biocompatibility and bioavailability owed to the lipid and lipid–PEG layers
In addition, the inner lipid layer also functions as a molecular fence that minimizes leakage of the
encapsulated content during the LPHNPs preparation
Furthermore, the inner lipid layer slows down the polymer degradation rate of the LPHNPs product by
limiting inward water diffusion, hence enabling sustained release kinetics of the content
Wide selections of biocompatible polymers and lipids and numerous polymer–lipid combinations
Easy fabrication by a single-step method
superior capability of co-encapsulating therapeutic and imaging agents of different properties
With such favourable characteristics, LPHNPs have rapidly evolved into a robust drug delivery platform
7
The LPHNPs exhibit
9. 9
The amount of drug in the outer shell and on
the particle surface is released in the form of
a burst
The chemical nature of the lipid is also
important because lipids which form highly
crystalline particles with a perfect lattice
(e.g. monoacid triglycerides) lead to drug
expulsion
Inherent low drug incorporation rate
Shows gelation tendency
High water content
PLNs show sustained drug release profile
Drugs are incorporated into the polymeric
core with high loading yields
Lipid monolayer at the interface of the
hydrophobic core and the hydrophilic shell
reduces water penetration
Disadvantages of SLNs Advantages of LPHNPs
ADVANTAGES OF LPHNPs OVER SOLID LIPID NANOPARTICLES
10. TYPES OF LPHNPs
LPHNPs
Monolithic or
Mixed lipid
polymer NPs
Biomimetic
LPH systems
Polymer-caged
liposomes
Core-shell
Polymer core–
lipid shell
Hollow core
lipid–polymer–
lipid systems
nanoparticles are coated
with RBC membrane
they are also known as
erythrocyte membrane-
camouflaged polymeric
nanoparticles
made up of polymeric
matrix in which the
lipid molecules are
dispersed throughout
When polymers are
anchored on the surface
of liposomes, polymer-
caged liposomes
are formed which are
more stable systems.
Such systems have a multi
layered structure comprising
of lipid-PEG and lipids in the
outermost layers which coats
the polymeric layer
surrounding a lipid layer with
inner most hollow core
11. Polymer core–lipid shell
Various terms like lipid polymer particle assemblies, lipid coated NPs, polymer
supported lipid shells, lipoparticles and nano-cell have been synonymously used for
these supramolecular lipid coated polymeric structures
The hybrid architecture made up of polymeric core and layers of lipid
provide several advantages
• Adjustable particle size and drug release
• ease of loading multiple agents
• better loading efficiency
• serum stability
11
13. METHOD OF PREPARATION FOR LPHNPs
SYNTHESIS APPROACH
Two step process Single step process
Polymer core and lipid shell prepared
separately and then merged together
Hybrid nanoparticles are prepared through
single step
10
15. 1. TWO STEP SYNTHESIS APPROACH
The polymeric core and lipid shell are prepared separately using two independent processes; then the
two components are combined by direct hydration, sonication, or extrusion to obtain the desired lipid
shell–polymer core structure
15
A. Conventional two-step method
16. STEP 1:
Preparation of polymeric nanoparticles
PNPs are prepared by emulsification solvent evaporation (EME), Nano-precipitation or
high pressure homogenization
STEP 2:
Preparation of lipid vesicles. The lipid component can be prepared in the form of a thin
dried film. Lipid is first dissolved in an organic solvent, like chloroform. Subsequently it is
subjected to rotary evaporation.
The next step is the hydration of this dried lipid film by solution of polymeric NPs
Lipid vesicles prepared by above mentioned method will be exposed to different mixing
protocols as given below
Vortexing: It is a low energy mixing process.
Ultra sonication 16
17. B. Non-conventional two-step method
polymeric nanoparticles (i.e., polyglutamic acid) (400–500 nm) were prepared by spray drying after
which they were dispersed in dichloromethane solution containing the lipids (i.e., tripalmitin,cetyl
alcohol)
The lipid–polymer suspension was later spray-dried to produce lipid-coated polymeric nanoparticles
17
Spray drying
Soft lithography particle molding technique
Particle Replication in Non-Wetting Templates (PRINT) was employed to prepare LPHNPs
for gene delivery
18. PRINT technology (Particle Replication in Non-Wetting Templates )
Polymer was dissolved in
an organic solvent (e.g
DMSO with the genetic
material (i.e siRNA)
Cast onto a polyethylene
terephthalate (PET) sheet
The PET sheet was then
heated while in conformal
contact with a PRINT
mold containing 80 * 320
nm patterns
allowing the polymer to
flow into the mold and to
solidify to ambient
temperature, thereby
forming the Polymer
nanoparticles
the nanoparticles were
harvested from the mold
by
having the mold in
conformal contact with a
poly(vinyl alcohol)(PVA)-
coated PET sheet
The nanoparticles were
then released from the
PVA-coated PET sheet
using aqueous solution of
the lipids (i.e DOTAP,
DOPE)
Freeze dried-needle
shaped
LPHNPs with length of
~200 nm
18
which dissolved the PVA layer, hence removing the
nanoparticles from the mold, and simultaneously forming
lipid-coated PLGA nanoparticles
20. Governing formulation parameters in the two-step method
size homogeneity of the preformed lipid vesicles
preformed vesicles prepared by extrusion were smaller and more uniform in size compared to vesicles
formed by thin lipid film hydration
Lipid formulation charge
the monodispersity of the formed LPNs also depended on the charge of the lipid vesicles
Minimal LPNs aggregation (i.e., narrow size distribution and high colloidal stability) was achieved by using
only one lipid type to form vesicles
lipid vesicle-to-polymeric nanoparticle ratio
vesicle to nanoparticle ratio (AV/AP) significantly influenced the LPNs’ colloidal stability
At high AV/AP and high DPTAP fractions-lipid vesicles acted as electrostatic stabilizers
At low AV/AP and low DPTAP fractions-Incomplete lipid coating of the nanoparticle core of one LPN led to the exposure
of its anionic surface to the cationic DPTAP of another LPN
20
AV/AP, and ionic strength of the continuous phase significantly
influence the resultant LPHNPs’ colloidal stability, and
consequently the size
21. 2. ONE STEP SYNTHESIS APPROACH
POLYMER LIPID HYBRID NANOPARTICLES
17
the stabilizing agents used, where the lipid functions as stabilizers of the LPNs produced, in place of the
ionic/non-ionic surfactants (e.g., PVA, Poloxamer) typically used in the non-hybrid polymeric nanoparticle
preparation
Lipid and lipid PEG conjugate self assemble on polymer nanoparticles
Polymer solution is added to lipid aqueous solution drop-wise so
polymer precipitate out
Lipid and lipid PEG conjugates in aqueous solution and co-solvent can
be added
Free polymer and hydrophobic drug in water miscible organic solvent
lipid monolayer is formed
hydrophobic tail of lipids- polymer core
hydrophilic head sticks -external aqueous
surrounding
resulting in the formation of LPNs that are stabilized by the
lipid.
One-step method by nanoprecipitation
Sonication
22. One-step method by emulsification–solvent–evaporation (ESE)
A single ESE method is employed when the substance to be encapsulated is soluble in a water-
immiscible solvent (i.e., oil phase)
the oil phase, which contains the polymer and the substance to be encapsulated, is added, under
constant stirring or ultrasonication, into an aqueous phase containing the lipid to form an oil-in-water
(o/w) emulsion.
When the oil phase is removed by evaporation, the polymer core is formed and simultaneously
the lipid self-assembles around the polymer– thus essentially forming the LPNs
22
ESE method
typically produces
larger LPNs
compared to the
nanoprecipitation
Single emulsification
23. employed when the substance to be encapsulated is insoluble in any organic solvents, such that it
cannot be dissolved together with the polymer
23
w/o emulsionw/o/w emulsion
evaporation of the oil phase,
gives rise to the LPNs
Double emulsification
24. RECENT ADVANCES IN ONE STEP METHOD OF NANO-PRECIPITATION
24(A) microfluidic nanoprecipitation to improve the LPHNPs size homogeneity
(B) multi-inlet vortex microreactor for high-throughput production of LPNs
25. FORMULATION PARAMETERS TO BE CONTROLLED
Lipid to Polymer Mass Ratio L/P Ratio
Higher L/P ratios led to concentrations higher than the critical micelle concentration resulting in the formation of
liposomes in addition to the LPNs, whereas lower L/P ratios led to LPNs aggregation due to insufficient lipid coating
The L/P ratio was also found to indirectly influence the encapsulation efficiency, loading, and release kinetics of the
encapsulated substance through its influence on the extent of the lipid coating of the polymer core
Lipid Coating
it acts as a molecular barrier that keeps the encapsulated substance inside the polymer core during the self-assembly
process results in High EE
lipid coating slows down the drug release kinetics by keeping the dissolution fluid medium away from the core
Lipid-PEG Fraction
Increasing the lipid PEG fraction resulted in more stable LPNs,
Type of Lipid
25
26. CHARACTERIZATION OF LPHNPs
26
Parameter Method of characterization
Physicochemical Properties
Particle size, PSD, Zeta Potential DLS (Zeta sizer), Photon correlation
spectroscopy(PCS) (10-150 nm)
Morphology SEM, TEM, CSLM, fluorescence microscopy
Drug loading
and entrapment
HPLC, dialysis , centrifugation, membrane filtration
Drug release
Stability study
In vivo evaluation
27. SURFACE FUNCTIONALISATION OF LPHNPs
PEG layer is essential to maintain stability of the hybrid nanoparticles :
In vitro by reducing nanoparticle aggregation
In vivo by allowing the particles to evade recognition by the reticuloendothelial system (RES)
and other immune cells
PEG molecules also provide functional groups for further modification of the hybrid nanoparticles
with targeting ligands for cell or tissue specific drug delivery
Targeting
ligands
Monoclonal
antibodies
Antibody
fragments
Aptamers Peptides Folic acid
18
32. SUMMARY
PLN system is a highly versatile drug delivery platform due to its diverse selection and combination of
polymer and lipid materials and highly modifiable nanostructure using these building blocks. Such
properties allow PLN to efficiently load single or multiple agents with vastly different physicochemical
properties
Superior efficacy and minimum tissue toxicity of PLN in pre-clinical studies suggest great potential of
LPHNPs for cancer treatments
Despite the great progress made on synthesis, characterization and applications of the hybrid
nanoparticles, we call attention to a few key unmet challenges in further developing this new
nanoparticle platform as a robust drug delivery
The simplicity of the synthesis process, especially the one-step self-assembly process, dramatically
increases the likelihood of producing the lipid-polymer hybrid nanoparticles in a scalable and
economical manner.
32
33. FUTURE DIRECTION
the multifunctional LPHNP platform with simultaneous imaging and drug delivery characteristics
could have important applications in cancer chemotherapy and diagnosis. Additionally, motivated by
the promises of gene therapy, there is much interest in developing LPHNP-mediated nonviral gene
delivery vectors for different therapeutic applications
recent developments in microfluidics and PRINT technology could increase the production scale of
LPHNPs in larger quantities with defined material characteristics
Despite these advances in the development and application of LPHNPs this field of research is in its
infancy and there are several challenges that need to be overcome to meet clinical expectations
We expect that LPHNPs will eventually replace current liposome and polymeric NP therapeutics
delivery systems
Future understanding of mechanisms that govern molecular interactions of PLN components and the
biological barriers will be very useful in designing multifunctional PLN systems for cancer treatment.
Clinical translation of PLN eventually for personalized nanomedicine could be distinctively possible.
33