This document presents information on aquasomes, a novel carrier for drug delivery. Aquasomes are spherical, nanoparticulate carrier systems composed of a solid nanocrystalline core coated with an oligomeric film. They are capable of stabilizing and delivering delicate biomolecules like proteins, peptides, hormones, and genes. The core provides structural stability while the carbohydrate coating protects molecules from degradation. Characterization techniques like TEM, SEM, and DSC are used to analyze aquasome size, structure, and glass transition temperature. Applications include insulin, enzyme, oxygen, antigen and drug delivery where aquasomes show improved stability and biological activity over unformulated drugs.

1. Presented to:
Dr. Shubhani Saraf
Head of Department
(Department of Pharmaceutical science)
Presented by:
Sishant Rav Divya
M.Pharm First Year ( Pharmaceutics )
BBAU Lucknow
Batch - 2017-19

2. Aquas mes

3. Contents..
o INTRODUCTION
o COMPOSITION OF AQUASOMES
o ROLE OF DISACCHARIDES IN AQUASOMES
o OBJECTIVES
o CHARECTERIZATION OF AQUASOMES
o PRINCIPLE OF SELF ASSEMBLY
o METHOD OF PREPARATION OF AQUASOME
o APPLICATION OF AQUASOME
o CONCLUSION
o REFERENCES

4. CARRIER FOR DRUG DELIVERY

5. AQUASOMES: A NOVEL CARRIER FOR DRUG DELIVERY
Drug
Core
C at

6. Introduction
• Recently developed delivery system for bioactive molecules
Such as
1. peptide
2. protein
3. hormones
4. enzymes
5. antigens and genes to specific sites
• It is like “bodies of water"
• These properties protect and preserve Delicate biological molecules

7. spherical in shape
60–300 nm particles size.
It is nanoparticulate carrier systems
but instead of nanoparticles 3 layered self assembled structures
Comprised a solid phase nanocrystalline
core coated with oligomeric film
biochemically active molecules adsorbed with or without modification
These structures self assembled by non-covalent & ionic bonds

8. • The solid core provides the structural stability
• while the carbohydrate coating protects against
Dehydration
• stabilizes the biochemically active molecules
• Aquasomes are first discovered by Nir Kossovsky.

9. COMPOSITION OF AQUASOMES
1- Core material
Ceramic and polymers are most widely used core materials.
Polymers such as
1. albumin
2. gelatin or acrylate
Ceramic such as
1. diamond particles
2. brushite (calcium phosphate)
3. tin oxide are used.

10. 2- Coating material
such as
• cellobiose, pyridoxal 5 phosphate, sucrose, trehalose, chitosan, citrate
etc.
• Carbohydrate act as natural stabilizer
3- Bioactive Molecules
• They have the property of interacting with film
• via non covalent and ionic interactions.

11. ROLE OF DISACCHARIDES IN AQUASOMES
• disaccharides contain a large quantity of hydroxyl group.
• help replace water around polar residues in proteins.
• Preserve aqueous structure of proteins on dehydration.
• OH- on oligomer interact with polar and charged groups of proteins.
maintaining integrity in absence of water.

12. OBJECTIVES

13. CHARECTERIZATION OF AQUASOMES
a. X-ray powder diffractometry
b. Transmission electron microscopy(TEM)
c. Scanning electron microscopy(SEM)
d. Drug Loading Efficiency
e. Particle size distribution
f. Zeta Potential
g. In-vitro drug release

14. Characterization of ceramic core
Characterization of ceramic core
Size distribution
a) scanning electron microscopy (SEM)
b) Transmission electron microscopy (TEM)
Core, coated core, drug-loaded aquasomes analyzed by these techniques.
particle size and zeta potential
a) photon correlation spectroscopy

15. Structural analysis
By using FT-IR spectroscopy
potassium bromide sample disk method
Core & coated core analyzed by recording their IR spectra
the wave number range 4000–400 cm–1

16. • Crystallinity
• crystalline or amorphous behavior using X-ray diffraction
• X-ray diffraction pattern of sample compared with standard diffractogram

17. Characterization of coated core
1. Carbohydrate coating
Induced aggregation method
(determines amount of sugar coated over core)
Anthrone method
(residual sugar unbound or residual sugar remaining after coating)
adsorption of sugar over the core by measurement of zeta potential

18. 2. Glass transition temperature
DSC (Differential Scanning Calorimetry)
• analyze effect of carbohydrate on drug to aquasomes
• study glass transition temperature of carbohydrates and proteins.
• transition from glass to rubber measured By DSC analyzer

19. Characterization of drug-loaded aquasomes
• Drug payload
incubating aquasome formulation (i.e. without drug)
In drug solution for 24 hours at 4°C
incubating aquasome formulation (i.e. without drug)
In drug solution for 24 hours at 4°C
centrifugation for 1 hour at low temperature
Drug remaining in the supernatant liquid after loading
Estimated by any suitable method of analysis

20. • In vitro release
Aquasomes incubating with drug-loaded
Samples are withdrawn periodically
Equal volumes of medium replaced after each withdrawal
supernatants analyzed for the amount of drug released

21. • In-process stability studies
• SDS-PAGE (sodium dodecyl sulphate polyacrylamide gel electrophoresis)
• Use to determine the stability and integrity of protein
• This process done during the formulation of aquasomes

22. PRINCIPLE OF SELF ASSEMBLY
1- Interaction between charged group
• such as amino,carboxyl, sulphate, phosphate groups
• Charged group plays role in stabilizing
tertiary structures of folded proteins.
2- Hydrogen bonding and dehydration effect
• H-bond helps in stabilize of secondary protein
such as alpha helices and beta sheets

23. 3- Structural stability
• Molecules have less charge than charged groups show dipole moment.
• stability of protein in biological environment determined by
- interaction between charged group and hydrogen bonds

24. Method of preparation of aquasomes
I- Formation of an inorganic core(ceramic core )
II- Carbohydrate coatings
III- Loading of the drug

25. I- Formation of an inorganic core(ceramic core )
1) Synthesis of nanocrystalline tin oxide core ceramic
tin oxide core ceramic can be synthesized
(by direct current sputtering)
high purity tin is sputtered in mixture of organ and oxygen gas.
ultra fine particles formed

26. 2) Self assembled nanocrystalline brushite (calcium phosphate)
solution of disodium hydrogen phosphate and calcium chloride
( by sonication OR colloidal precipitation)
Self assembled nanocrystalline
2Na2HPO4 + 3CaCl2 + H2O → Ca3 (PO4)2 + 4NaCl+ 2H2 + Cl2 + (O)

27. 3) Nanocrystalline carbon ceramic, diamond particles
for the core synthesis
after ultra cleansing and sonication
The size range between 50-150nm
It exibit reactive surface.

28. II-Carbohydrate coatings
ceramic cores are coated with carbohydrate (polyhydroxyl oligomer)
addition of carbohydrate into aqueous dispersion of cores under sonication
lyophilization to promote an irreversible adsorption of carbohydrate into the ceramic surface
unadsorbed carbohydrate is removed by centrifugation

29. Example: of coating materials are
1. Cellobiose
2. Citrate
3. Pyridoxal-5- phosphate
4. Trehalose
5. Sucrose.etc

30. III- Loading of the drug of choice to this assembly
solution of drug is prepared in suitable pH buffer
obtain the drug-loaded formulation
coated particles are dispersed into it

31. Method of preparation of aquasomes

34. Applications of aquasomes
• I- Insulin delivery
• II- Oral delivery of acid labile enzyme
• III- As oxygen carrier
• IV- Antigen delivery
• V- Delivery of drug
• VI- For delivery of gene

35. I- Insulin delivery
• Cherian et al prepared
calcium phosphate ceramic core for parenteral delivery of insulin
core was coated with cellobiose,trehalose, and pyridoxal-5-phosphate
aquasome formulations of insulin was evaluated using albino rats.
Prolonged reduction of blood glucose was observed

36. II- Oral delivery of acid labile enzyme
• Rawat et al Proposed
Nanocore was prepared by colloidal precipitation under sonication at room temperature
core was then coated with chitosan
enzyme was adsorbed
enzyme was protected by alginate gel.
in vitro drug release data
aquasomes were found to be protecting structural integrity of enzymes
to obtain a better therapeutic effect

37. III- As oxygen carrier
• Khopade et al prepared
hydroxyapatite core by using carboxylic acid
Core was coated with trehalose
adsorption of haemoglobin (Hb)
loading capacity was found to be approx. 13.7 mg of haemoglobin
Studies carried out in rats showed good potential for oxygen carrier

38. In another study
• Patil and co-workers prepared
Hydroxyapatite ceramic cores by co-precipitation and self-precipitation
coated with various sugars maltose, and sucrose
(Hb)hemoglobin was adsorbed
Drug loading was estimated by the benzidine method.
Oxygen carrying capacity was found to be similar to fresh blood

39. IV- Antigen delivery (200 nm)
• Vyas et al prepared
Hydroxyapatite ceramic cores by co-precipitation method
coated with cellobiose and trehalose,
bovine serum albumin was adsorbed
The antigen-loading efficiency was found to be approximately 20–30%.
formulation was compared to plain bovine serum albumin,
was found to show a better response

40. V- Delivery of drug (60–120 nm)
• Oviedo and co-workers prepared
Aquasome core of calcium phosphate
covered with a lactose film
adsorption of Indomethacin as a low-solubility drug
aquasomes were characterized
Particle size was found to be in the range of 60–120 nm.
release studies from these carriers are yet to be determined

41. VI- For delivery of gene
Aquasomes used for successful targeted intracellular gene therapy
A five layered composition comprised of the ceramic nanocrystalline core,
polyhydroxyl oligomeric film coating
the non covalently bound layer of therapeutic gene segment,
additional carbohydrate film coating
targeting layer conserved viral membrane proteins

43. CONCLUSION
• The drug candidates delivered through the aquasomes show better
biological activity due to the presence of the unique carbohydrate
coating the ceramic. the crystalline nature of the core gives structural
stability and overall integrity

44. REFERENCES
• 1. N. Kossovsky, A. Gelman, E.E. Sponsler, H.J. Hnatyszyn, S. Rajguru, M.
Torres, et al., Biomaterials, 1994; 15, 1201.
• 2. Arakawa, T. and Timasheff, S. N.”Stabilization of protein structure by
sugars.” Biochemistry, 1982; 21:6536-6544.
• 3. Crowe, J.H.; Crowe, L.M.; Carpenter, J.F.; Rudolph, A.S.; Wistrom. C.A;
Spargo, B.J. and Acnhordoguy. T.J ”Interaction of sugars with membrane”.
Biochem biophys acta 1947:367-384.
• 4. Kossovsky, Gelman.A. and Sponsler, E.E. ”Cross linking encapsulated
haemoglobin solid phase supports: lipid enveloped haemoglobin adsorbed
to surface modified ceramic particles exhibit physiological oxygen lability
artif.cells blood sub”biotech, 1994; 223: 479-485

45. • 21. S.M. Moghimi, A.C. Hunter, and J.C. Murray. Nanomedicine: current
status and future prospects. FASEB J. 2005; 19: 311-330.
• 22. R.D. Handy, F.D. Kammer, J.R. Lead, M. Hassello¨v, R. Owen, M.
Crane, Ecotoxicology, 2008; 17, 287.
• 23. Irma Rojas-Oviedo, Rodrigo A. Salazar-L ´opez, “Elaboration and
structural analysis of aquasomes loaded with Indomethacin” european
journal of pharmaceutical sciences Nov; 1994; 32(3):223-30.
• 24. Kossovsky, N.”Perfecting delivery”chemistry in Britain. 43-45. 1996.
• 25. Kossovsky, N.; Millet, d; Gelman, L.A.; Sponsler.E.Dand
Hnatyszyn.H.J”Self assembling nano structures” matr. Res. Soc bull. Seot.
1993. 78-81.