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KOSARAJU SAI VIVEK
I M.PHARMACY,
DEPARTMENT OF
PHARMACEUTICS
JSS COLLEGE OF
PHARMACY
AQUASOMES
 Aquasomes are first discovered by Nir Kossovsky.
 These are nanoparticulate carrier systems with
three layered self assembled structures
These comprises of central solid nanocrystalline core
coated with polyhydroxy oligomers onto
which biochemically active molecules are adsorbed.
 Aquasomes are also called as “bodies of water” and
their water like properties protect and preserve fragile
biological molecules,
 This property of maintaining conformational
integrity as well as high degree of surface exposure
made it as a successful carrier system for bioactive
molecules like peptide, protein, hormones, antigens
and genes to specific sites, that is for targeting.
PROPERTIES
1. Aquasomes water like properties provides a platform for
preserving the conformational Integrity and bio chemical
stability of bio-actives.
2. Aquasomes due to their size and structure stability, avoid
clearance by reticuloendothelial system or degradation by
other environmental challenges
3. Aquasomes possess large size and active surface hence can
be efficiently loaded with substantial amounts of agents
through ionic, non covalent bonds, van der waals forces and
entropic forces. As solid particles dispersed in aqueous
environment, they exhibit physical properties of colloids
 In general these aquasomes are assemblies of
simple polymers, complex lipid mixtures with
diameter ranging between 30 to 500 nm.
 As these are solid or glassy particles dispersed in
an aqueous environment, they exhibit the
physical properties of colloids and their mechanism
of action is controlled by their surface chemistry.
 Aquasomes deliver their contents through a
combination of specific targeting, slow and
sustained release process
METHOD OF PREPARATION OF
AQUASOMES:
 By using the principle of self assembly, the
aquasomes are prepared in three steps i.e.,
1. preparation of core,
2. coating of core, and
3. immobilization of drug molecule.
Principle of self assembly
 In aqueous biological environments , the assembly of
macro molecule Is governed by three process.
 (1) Interaction between charged group.
(2) Hydrogen bonding and dehydration effect.
(3) structural stability.
1) Interaction between charged
group
 Most of the Biological product are charged due to intrinsic
chemical group or absorbed ion from the biological
environment.
 Interaction of charged group such as
amino, carbonyl, sulphate, phosphate groups facilitate the
long range approach of self assembling sub units.
 Charged groups also play role in stabilizing tertiary structure
of folded proteins.
 Example of ion pairs -carboxylated /phosphate group bound
to ionized arginine / lysine side chain of protein.
2)Hydrogen bonding and dehydration
effect.
 Hydrogen bond are formed between hydrogen atom
attached to an electronegative donor atom (Ex
oxygen ,Nitrogen,) and an electronegative or basic
acceptor (Ex carbonyl oxygen).
 Hydrogen bond help in base pair matching and
stabilization of Secondary protein structure.
 Molecule that form hydrogen bonds are hydrophilic
and these molecules confer significant degree of
organization to the surrounding water molecules.
3) structural stability.
 The structural stability of Protein in the biological
environment is determined by the interaction
between charged groups and hydrogen bond largely
external to the molecule and vander walls forces
largely internal to the molecule.
 Vander walls forces are largely responsible for the
hardness or softness of the molecule. The vander
walls interaction among hydrophilic side chains
promotes stability of compact helical structures.
 Method of preparation :
 By using the principle of self assembly Aquasomes
can be prepared by three method.
(1) Preparation of core.
(2) coating of core.
(3) Immobilization of drug molecule.
1) Preparation of core.
 This stage mainly depends on the
selection of material for core.
-its physical chemical properties
 ‡ This can be fabricated by the
-Sonication
-Colloidal precipitation..
 For the core material material ceramic material widely
used ,as they are structurally to be known..
 Commonly used ceramic core are tin oxide, and
calcium phosphate.
 Example:
 synthesis of nanocrystalline tin oxide core material.
 This can be prepared by
-Direct current reactive.
-Magnetron sputtering.
3 inch diameter target of highly purified Tin is sputtered
in
High pressure gas mixture of argon and oxygen.
The ultra fine particle form in gas phase are collect on
copper tube and cool at 70oK with liquid nitrogen
Synthesis of nano crystalbrushite
(calciumphosphate dihydrate)
 ‡This can be prepared by
-colloidal dispersion
-Sonication
-By reaction of disodium hydrogen phosphate
and calcium phosphate.
 The commonly feature include
-Crystalline
-They measure b/w 50-150nm. And exhibit clean
and reactive surface
(2) Carbohydrate coating
 The second step involves coating by carbohydrate
on the surface of ceramic cores.
 There are number of processes to enable the
carbohydrate (polyhydroxy oligomers) coating to
adsorb epitaxially on to the surface of the Nano
crystalline ceramic cores.
Process generally entail,
 Addition of poly hydroxy oligomer,
To a dispersion of core in ultra pure water.
Lyophilization (to promote the adsorption of
carbohydrate on the surface
of ceramic core)
Excess of carbohydrate is removed by
stir cell ultrafilteration.
 Commonly used coating material,
 - Cellobiose
- Citrate
- Sucrose
- Trihalose
- Pyridoxal -5- phosphate.
 (3) Immobilization of drug
 The surface modified Nano crystalline core provide the solid
phase for subsequent non denaturing self assembly for a broad
range of biological active molecule.
 Drug can be loaded by partial adsorption
Characterization of ceramic core
 Size distribution:
 For morphological characterization and size
distribution analysis, scanning electron
microscopy (SEM) and transmission electron
microscopy (TEM) are generally used.
 Core, coated core, as well as drug-loaded aquasomes
are analyzed by these techniques.
 Mean particle size and zeta potential of the particles
can also be determined by using photon correlation
spectroscopy
 Structural analysis:
1. FT-IR spectroscopy can be used for structural analysis. Using
the potassium bromide sample disk method, the core as well
as the coated core can be analyzed by recording their IR
spectra in the wave number range 4000-400 cm-1;
2. The characteristic peaks observed are then matched with
reference peaks. Identification of sugar and drug loaded
over the ceramic core can also be confirmed by FT-IR
analysis of the sample.
 Crystallinity:
The prepared ceramic core can be analyzed for its crystalline or
amorphous behavior using X-ray diffraction. In this
technique, the X-ray diffraction pattern of the sample is
compared with the standard diffractogram, based on which the
interpretations are made.
Characterization of coated core
 Carbohydrate coating
 Coating of sugar over the ceramic core can be
confirmed by
i. concanavalin A-induced aggregation method
(determines the amount of sugar coated over core) or
ii. anthrone method (Determines the residual sugar
unbound or residual sugar remaining after coating).
iii. Furthermore, the adsorption of sugar over the core
can also be confirmed by measurement of zeta
potential
Characterization of drug-loaded
aquasomes
 Drug payload
i. The drug loading can be determined by measuring the drug
remaining in the supernatant liquid after loading which can
be estimated by any suitable method of analysis 19.
 In vitro drug release studies
i. The in vitro release kinetics of the loaded drug is determined
to study the release pattern of drug from the aquasomes by
incubating a known quantity of drug-loaded aquasomes in a
buffer of suitable pH at 37 °C with continuous stirring.
ii. Samples are withdrawn periodically and centrifuged at high
speed for certain lengths of time. Equal volumes of medium
must be replaced after each withdrawal. The supernatants are
then analyzed for the amount of drug released by any
suitable method .
APPLICATIONS OF AQUASOMES
 Aquasomes has got a quite versatile application potential
as a carrier for delivery of
vaccines, hemoglobin, drugs, dyes, enzymes.
1) Aquasomes used as vaccines for delivery of viral antigen
2) Aquasomes as red blood cell substitutes can effectively
deliver the large, complex labile molecule, haemoglobin. By
incorporating in aquasome carriers, the toxicity of
haemoglobin is reduced, biological activity is preserved,
haemoglobin concentration of 80% can be achieved and is
reported to deliver oxygen in a non linear manner like
natural red blood cells.
3) Aquasomes for pharmaceuticals delivery i.e.
insulin, developed because drug activity is
conformationally specific. Bio activity preserved and
activity increased to 60% as compared to i.v.
administration and toxicity not reported .
4) Aquasomes are used for oral delivery of acid labile
enzyme, serratiopeptidase. Enzyme loaded aquasome was
further protected by encapsulating in alginate gel.
They protected structural integrity of enzymes and
better therapeutic efficacy was observed .

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Aquasomes

  • 1. KOSARAJU SAI VIVEK I M.PHARMACY, DEPARTMENT OF PHARMACEUTICS JSS COLLEGE OF PHARMACY AQUASOMES
  • 2.  Aquasomes are first discovered by Nir Kossovsky.  These are nanoparticulate carrier systems with three layered self assembled structures These comprises of central solid nanocrystalline core coated with polyhydroxy oligomers onto which biochemically active molecules are adsorbed.
  • 3.
  • 4.  Aquasomes are also called as “bodies of water” and their water like properties protect and preserve fragile biological molecules,  This property of maintaining conformational integrity as well as high degree of surface exposure made it as a successful carrier system for bioactive molecules like peptide, protein, hormones, antigens and genes to specific sites, that is for targeting.
  • 5. PROPERTIES 1. Aquasomes water like properties provides a platform for preserving the conformational Integrity and bio chemical stability of bio-actives. 2. Aquasomes due to their size and structure stability, avoid clearance by reticuloendothelial system or degradation by other environmental challenges 3. Aquasomes possess large size and active surface hence can be efficiently loaded with substantial amounts of agents through ionic, non covalent bonds, van der waals forces and entropic forces. As solid particles dispersed in aqueous environment, they exhibit physical properties of colloids
  • 6.  In general these aquasomes are assemblies of simple polymers, complex lipid mixtures with diameter ranging between 30 to 500 nm.  As these are solid or glassy particles dispersed in an aqueous environment, they exhibit the physical properties of colloids and their mechanism of action is controlled by their surface chemistry.  Aquasomes deliver their contents through a combination of specific targeting, slow and sustained release process
  • 7. METHOD OF PREPARATION OF AQUASOMES:  By using the principle of self assembly, the aquasomes are prepared in three steps i.e., 1. preparation of core, 2. coating of core, and 3. immobilization of drug molecule.
  • 8. Principle of self assembly  In aqueous biological environments , the assembly of macro molecule Is governed by three process.  (1) Interaction between charged group. (2) Hydrogen bonding and dehydration effect. (3) structural stability.
  • 9. 1) Interaction between charged group  Most of the Biological product are charged due to intrinsic chemical group or absorbed ion from the biological environment.  Interaction of charged group such as amino, carbonyl, sulphate, phosphate groups facilitate the long range approach of self assembling sub units.  Charged groups also play role in stabilizing tertiary structure of folded proteins.  Example of ion pairs -carboxylated /phosphate group bound to ionized arginine / lysine side chain of protein.
  • 10. 2)Hydrogen bonding and dehydration effect.  Hydrogen bond are formed between hydrogen atom attached to an electronegative donor atom (Ex oxygen ,Nitrogen,) and an electronegative or basic acceptor (Ex carbonyl oxygen).  Hydrogen bond help in base pair matching and stabilization of Secondary protein structure.  Molecule that form hydrogen bonds are hydrophilic and these molecules confer significant degree of organization to the surrounding water molecules.
  • 11. 3) structural stability.  The structural stability of Protein in the biological environment is determined by the interaction between charged groups and hydrogen bond largely external to the molecule and vander walls forces largely internal to the molecule.  Vander walls forces are largely responsible for the hardness or softness of the molecule. The vander walls interaction among hydrophilic side chains promotes stability of compact helical structures.
  • 12.  Method of preparation :  By using the principle of self assembly Aquasomes can be prepared by three method. (1) Preparation of core. (2) coating of core. (3) Immobilization of drug molecule.
  • 13. 1) Preparation of core.  This stage mainly depends on the selection of material for core. -its physical chemical properties  ‡ This can be fabricated by the -Sonication -Colloidal precipitation..  For the core material material ceramic material widely used ,as they are structurally to be known..
  • 14.  Commonly used ceramic core are tin oxide, and calcium phosphate.  Example:  synthesis of nanocrystalline tin oxide core material.  This can be prepared by -Direct current reactive. -Magnetron sputtering.
  • 15. 3 inch diameter target of highly purified Tin is sputtered in High pressure gas mixture of argon and oxygen. The ultra fine particle form in gas phase are collect on copper tube and cool at 70oK with liquid nitrogen
  • 16. Synthesis of nano crystalbrushite (calciumphosphate dihydrate)  ‡This can be prepared by -colloidal dispersion -Sonication -By reaction of disodium hydrogen phosphate and calcium phosphate.  The commonly feature include -Crystalline -They measure b/w 50-150nm. And exhibit clean and reactive surface
  • 17. (2) Carbohydrate coating  The second step involves coating by carbohydrate on the surface of ceramic cores.  There are number of processes to enable the carbohydrate (polyhydroxy oligomers) coating to adsorb epitaxially on to the surface of the Nano crystalline ceramic cores.
  • 18. Process generally entail,  Addition of poly hydroxy oligomer, To a dispersion of core in ultra pure water. Lyophilization (to promote the adsorption of carbohydrate on the surface of ceramic core) Excess of carbohydrate is removed by stir cell ultrafilteration.
  • 19.  Commonly used coating material,  - Cellobiose - Citrate - Sucrose - Trihalose - Pyridoxal -5- phosphate.  (3) Immobilization of drug  The surface modified Nano crystalline core provide the solid phase for subsequent non denaturing self assembly for a broad range of biological active molecule.  Drug can be loaded by partial adsorption
  • 20.
  • 21. Characterization of ceramic core  Size distribution:  For morphological characterization and size distribution analysis, scanning electron microscopy (SEM) and transmission electron microscopy (TEM) are generally used.  Core, coated core, as well as drug-loaded aquasomes are analyzed by these techniques.  Mean particle size and zeta potential of the particles can also be determined by using photon correlation spectroscopy
  • 22.  Structural analysis: 1. FT-IR spectroscopy can be used for structural analysis. Using the potassium bromide sample disk method, the core as well as the coated core can be analyzed by recording their IR spectra in the wave number range 4000-400 cm-1; 2. The characteristic peaks observed are then matched with reference peaks. Identification of sugar and drug loaded over the ceramic core can also be confirmed by FT-IR analysis of the sample.  Crystallinity: The prepared ceramic core can be analyzed for its crystalline or amorphous behavior using X-ray diffraction. In this technique, the X-ray diffraction pattern of the sample is compared with the standard diffractogram, based on which the interpretations are made.
  • 23. Characterization of coated core  Carbohydrate coating  Coating of sugar over the ceramic core can be confirmed by i. concanavalin A-induced aggregation method (determines the amount of sugar coated over core) or ii. anthrone method (Determines the residual sugar unbound or residual sugar remaining after coating). iii. Furthermore, the adsorption of sugar over the core can also be confirmed by measurement of zeta potential
  • 24. Characterization of drug-loaded aquasomes  Drug payload i. The drug loading can be determined by measuring the drug remaining in the supernatant liquid after loading which can be estimated by any suitable method of analysis 19.  In vitro drug release studies i. The in vitro release kinetics of the loaded drug is determined to study the release pattern of drug from the aquasomes by incubating a known quantity of drug-loaded aquasomes in a buffer of suitable pH at 37 °C with continuous stirring. ii. Samples are withdrawn periodically and centrifuged at high speed for certain lengths of time. Equal volumes of medium must be replaced after each withdrawal. The supernatants are then analyzed for the amount of drug released by any suitable method .
  • 25. APPLICATIONS OF AQUASOMES  Aquasomes has got a quite versatile application potential as a carrier for delivery of vaccines, hemoglobin, drugs, dyes, enzymes. 1) Aquasomes used as vaccines for delivery of viral antigen 2) Aquasomes as red blood cell substitutes can effectively deliver the large, complex labile molecule, haemoglobin. By incorporating in aquasome carriers, the toxicity of haemoglobin is reduced, biological activity is preserved, haemoglobin concentration of 80% can be achieved and is reported to deliver oxygen in a non linear manner like natural red blood cells.
  • 26. 3) Aquasomes for pharmaceuticals delivery i.e. insulin, developed because drug activity is conformationally specific. Bio activity preserved and activity increased to 60% as compared to i.v. administration and toxicity not reported . 4) Aquasomes are used for oral delivery of acid labile enzyme, serratiopeptidase. Enzyme loaded aquasome was further protected by encapsulating in alginate gel. They protected structural integrity of enzymes and better therapeutic efficacy was observed .