Aquasomes: A Nanoparticulate approach for the Delivery of Drug

Prathamesh L. Shirole, Dhanashree P. Sanap*, Kisan R. Jadhav

Bharati Vidyapeeth’s College of Pharmacy, Sector-8, CBD Belapur, Navi Mumbai 400614, Maharashtra, India.

*Corresponding Author E-mail: dhanashree.sanap@bvcop.in

ABSTRACT:

Aquasomes are one of the most recently developed delivery systems that are found a niche as peptide and protein carriers. Aquasomes are recently developed delivery systems for bioactive molecules like peptides, proteins, hormones, antigens, and genes to specific sites. The delivery system has been successfully utilized for the delivery of insulin, hemoglobin, and enzymes like serratiopeptidase, etc. Aquasomes are nanoparticles made up of calcium phosphate or ceramic diamond coated with a polyhydroxyoligomeric film. They are round nanoparticulate drug delivery systems of around 60-300 nm in size with three layered self-assembled structures. The solid core provides structural stability, while the carbohydrate coating protects against dehydration and stabilizes the biochemically active molecules. Aquasomes can also be referred to as "bodies of water". These water-like characteristics of aquasomes shield and retain delicate biological molecules. Aquasomes are prepared in three steps such as the development of the core, coating of the core, and the loading of the drug. Enzyme activity and sensitivity towards molecular conformation made aquasome a novel carrier for enzymes like DNases and pigment/dyes. The market potential and prospects of aquasomes are given in the article. This review article also includes a brief introduction to aquasomes, method of preparation, advantages, and disadvantages, evaluation parameters, and applications of aquasomes.

KEYWORDS: Aquasomes, Advantages, Nanoparticles, Ceramics, Applications.

INTRODUCTION:

Nir Kossovsky invented aquasome, a self-assembling nanoparticulate carrier system, in 1995. Kossovsky invented a method for creating nanoparticles carrying aquasomes, which are suitable for parenteral delivery due to their small particle size (less than 1000 nm), which eliminates the clogging of blood capillaries^1,2. Aquasomes have so been thoroughly researched for the delivery of medications with both tiny and large molecular weights. The primary benefit of aquasomes over other nanoparticulate carrier systems is that there is no drug-carrier interaction³.

Aquasome has a ceramic core that has been coated with a polyhydroxy oligomer, and on top of that biochemically active molecules are introduced through co-polymerization, diffusion, or adsorption.

Bones are good biomaterial for use as a drug carrier, since they have ideal biocompatibility, biodegradability, lack of toxicity, and stability due to the presence of calcium phosphate. Hydroxyapatite and calcium phosphate can be used as ceramic core materials in aquasomes formulation. Aquasomes often form by ionic interactions, non-covalent bonding, and vander waals forces⁴.

Aquasomes discovery comprises a principle from microbiology, food chemistry, biophysics and many discoveries including solid phase synthesis, supramolecular chemistry, molecular shape change and self-assembly⁵. Aquasomes can also be referred to as "bodies of water". This water-like characteristics of aquasomes shield and retain delicate biological molecules⁶. The surface chemistry of aquasomes regulates how they function. Aquasomes distribute their contents by combining precise targeting, molecular shielding, and a slow, prolonged release process. Bioactive molecules like peptide and protein hormones, enzymes, antigens, and genes are utilised in the targeting of molecules to certain places because they have the ability to preserve conformational integrity and have a high degree of surface exposure⁵.

METHOD OF PREPARATION:

There are three steps in the aquasomes preparation. The general process entails:

1. Development of an inorganic core

2. Polyhydroxy oligomer coating of the core and

3. Loading with the desired medication

The development of an inorganic core Common ceramic core materials include calcium phosphate, hydroxyapatite and diamond. These materials are used for the development of the core using colloidal precipitation and sonication method. The precipitated cores are separated by centrifugation and the sodium chloride that was produced during the reaction is removed by washing with enough distilled water. To get the particles of a particular size, the precipitates are then re-suspended in distilled water and filtered through a fine membrane filter^7,8.

4Na₂HPO₄ + 3CaCl₂⟶Ca₃ (PO₄)₂ + 2NaH₂PO₄ + 6NaCl

Figure 1: Preparation of aquasome⁷

Polyhydroxy oligomer coating of the core: Polyhydroxy oligomer coating can be achieved by adding carbohydrates to an aqueous dispersion of the cores under sonication. To encourage irreversible adsorption of carbohydrates onto the ceramic surface, it is also lyophilized. Adsorption, non-solvent addition and direct incubation methods can also be used to coat a core. Centrifugation is used to separate the unadsorbable carbohydrates. Cellobiose, citrate, pyridoxal-5-phosphate, trehalose and sucrose are some of the coating substances that are frequently used^2,9.

Loading with desired medication: Finally, adsorption loads the medication onto the coated particles. For that, coated particles are dispersed into a solution of known drug concentration made in a suitable pH buffer. The dispersion is then either lyophilized after some time to produce the drug-loaded formulation or stored overnight at a low temperature for drug loading (i.e aquasomes). The preparation is then characterized by utilizing a variety of methods¹⁰.

Advantages and Disadvantages of Aquasomes:

Aquasome functions as a reservoir to release the drug molecules either continuously or pulsatilely, reducing the need for multiple injections.

Figure 2: Advantages and Disadvantages of Aquasome⁷

Recent Patents:

Table 1: Recent Patents on Aquasomes

Patent no

Title

Inventors

Applicant

Claims

Composition

Method of preparation

Ref

PCT/IN2021/050832

Polyhydroxy oligomer coated dolutegravir aquasomes and method thereof

Pavani Govada,

Mastanamma SK

Varalakshmi Mummidi

The ratio of core: sugar coating: drug is 3:3-6:1

It is composed of inorganic core of calcium phosphate, carbohydrates, or polyhydroxy oligomers and the drug.

i. Ceramic core was prepared ii. Sugar coating was done on ceramic core

iii. And drug was adsorption on the coated ceramic

16/221,540

Gel formulation to treat diabetic foot ulcer infections

Marakanam Srinivasan, Uma Shankar

SRM Institute of Science and Technology

An antibiotic-loaded aquasomes

A gelling agent, a preservative, and a fluid medium

Formation of antibiotic-loaded aquasomes and then blending of loaded aquasomes

Novel Technologies/Various Investigations:

Table 2: Various investigation on aquasomes as vesicular drug delivery system¹³

Sr No	Formulators	Drug	Experiment
1	Nanjwade et al., 2013	Etoposide	Etoposide aquasomes were created by adsorbing the etoposide to a calcium phosphate inorganic core that was then wrapped with a lactose film. It was discovered that the diameter of aquasomes containing drugs ranged from 150 to 250 nm. Entrapment effectiveness was discovered to be 88.41%. Drug-loaded nanoparticles' average targeting efficiency was 42.54 % of the injected dose in the liver, 12.22 % in the lungs, 4.14 % in the kidney and 25.12 % in the spleen. Nanoparticles demonstrated superior drug targeting to the liver, spleen, lungs and kidney, according to the results.
2	Vengal et al., 2013	Piroxicam	To investigate the connection between particle size and dissolution profile, piroxicam, a drug that is poorly soluble in water, was synthesized as ceramic nanoparticles. The ceramic nanoparticles' yield was 66.7 %. It was noted that piroxicam was released from the ceramic nanoparticle formulations after 1 hour and 15 minutes.
3	Tiwari et al., 2012	Dithranol	Aquasomes were created using the colloidal precipitation method and added to a lotion to treat psoriasis. It was discovered that the drug-loading efficiency was 84.8 % w/w. In 7 hours, a 55.93 % drug release was seen. Aquasome-loaded cream controlled the drug release in comparison to plain cream, according to in vitro drug release experiments from both creams.
4	Kommineni et al., 2012	Insulin	The typical procedure used for aquasomes was used to formulate insulin-containing aquasomes. When standard pig insulin solution was paired with the framed aquasome's in vivo data, positive outcomes were achieved in contrast to insulin solution.
5	Cherian et al., 2000	Piroxicam	By using refluxing, sonication and coprecipitation, ceramic nanoparticles were created. The sonication method of core preparation was chosen because of its higher yield (42.4 ±0.4%) and shorter time requirement (1 day) compared to the reflux approach (27.4 ± 2.05%, 6 days). For the lactose-coated core and the piroxicam-loaded aquasomes, the morphological analysis showed spherical nanoparticles with sizes of 56.56 nm, 5.93 nm and 184.75nm, 13.78 nm, respectively, validating the nanometric dimensions.

Evaluation Parameters:

Size and shape:

After negative staining with a phosphotungstic acid solution, aquasomes can be examined morphologically under a transmission electron microscope. The Autosizer II C instrument and SEM are used in a photon correlation spectroscopy to estimate the mean particle size and size distribution. The key characteristics of aquasomes are their shape, particle size and structural analysis. Through x-ray powder diffractometry, the chemical makeup and crystalline structure of all samples were determined^14,15.

Structure Analysis:

The KBr pellet method is used for the structural investigation using fourier transform infrared spectroscopy (FTIR) in the wave number ranging from 400-4000 cm^-1. By using this technology, ceramic core, carbohydrate-coated core, drug-loaded formulation, and drug are all examined. FTIR analysis can be used to determine the drug's stability in its formulation¹.

Glass transition temperature:

Differential scanning calorimetry (DSC), which has been used to research the glass transition temperature of proteins and carbs, can be used to examine the impact of carbohydrates on the medication loaded into aquasomes. When the glass melts, the shift from the glass to the rubber state can be approximated as a temperature change⁷.

Crystallinity:

To determine the crystalline behavior of the ceramic core, x-ray diffraction is performed¹⁶.

Carbohydrate Coating:

The amount of sugar coating on the ceramic core can be determined using the concanavalin A-induced aggregation method or the anthrone method (determines the residual sugar unbound or residual sugar remaining after coating). Additionally, measurement of zeta potential can be used to establish the adsorption of sugar across the core^17,18.

Drug loading efficiency:

This test is carried out to verify the quantity of drug loaded in the aquasomes' formulation. Utilizing solutions of 0.1N methanolic hydrochloric acid, hydrophobic drugs like indomethacin and piroxicam is evaluated by subjecting it to spectroscopic examination¹⁹.

In vitro drug release studies:

By incubating a known quantity of drug-loaded aquasomes in a buffer with a suitable pH at 37°C with continuous stirring, the in vitro release kinetics of the loaded drug is evaluated to examine the release pattern of the drug from the aquasomes. Samples are periodically removed and quickly centrifuged for predefined lengths of time. After each withdrawal, equal volume of the medium must be replaced. The amount of medication released from the supernatants is then determined using any appropriate method¹⁴.

Zeta Potential Measurement:

The electrostatic attraction or repulsion between particles is measured by the zeta potential. It is the most reliable sign of dispersions stability, including suspension and emulsion. The intensity and makeup of an aquasome's surface charge are crucial because they determine how the organism will interact with its surrounding biologically and through electrostatic interactions with bioactive compounds^9,20. The kind of carbohydrate (oligomer) utilized to make aquasomes affects the zeta potential value. In a prior investigation, it was discovered that the zeta potential values of aquasomes made from trehalose, cellobiose, and pyridoxal-5-phosphate were, respectively, 15.6 1.15 mV, 20.4 0.9 mV, and 23.2 1.26 mV. It can also be used to confirm that sugar has been absorbed over the core. It was discovered that an increase in the carbohydrate saturation process on the hydroxyapatite core causes a decrease in zeta potential value^7,21.

Applications of Aquasomes:

Oral delivery of enzyme:

Serratiopeptidase, an acid-labile enzyme, is administered orally using a nanosized ceramic core-based approach. By using colloidal precipitation and sonication at room temperature, the nanocore was created. The chitosan was enforced to the core and continuously stirred before the enzyme was adsorbed on top of it. It was discovered that these aquasomes were preserving the structural integrity of enzymes to improve the therapeutic effect^8,22. DNase activity fluctuates in polypeptides like an enzyme with changes in molecular shape, but the enzyme's bioactivity is maintained. The long-term toxicity is not characterized¹⁹.

Vaccine Delivery:

The development of the subunit vaccine has stalled due to its low immunogenicity and stability. However, Goyal et al., (2006) developed aquasomes that contained hepatitis B surface antigen (HBsAg)²³. When the HBsAg was put onto the cellobiose-coated hydroxy apatite core, its size and spherical shape were nanometric. Because of this, aquasome is a desirable approach for administrating vaccines^9,24.

Delivery of Gene:

Targeted intracellular gene therapy has been successfully carried out using aquasomes, which feature five layers: a ceramic core, a polyoxyoligomeric film, a therapeutic gene segment, an extra carbohydrate film, and a targeting layer of a conformationally conserved viral membrane protein²⁵.

Figure 3: Gene delivery through aquasomes

Insulin Delivery:

Drug activity is conformationally specific, leading to the development of aquasomes for the administration of pharmaceuticals like insulin. Compared to intravenous treatment, bioactivity was conserved, activity rose to 60%, and toxicity was unreported²⁶.

Anti-thrombic activity:

Because the release of oxygen by haemoglobin is conformationally sensitive, aquasomes serve as substitutes for red blood cells with haemoglobin bound on oligomer surfaces. This lowers toxicity, achieves an 80% haemoglobin content, and is reported to deliver blood nonlinearly like normal blood cells²⁵.

Figure 4: Anti-thrombic activity of aquasomes⁹

Antigen Delivery:

When Epstein-Barr and immune deficiency viruses are delivered via aquasomes as vaccines, the goal of vaccination therapy must be triggered by conformationally specific target molecules to elicit the proper antibodies²¹. Antibodies against a conformationally specific target mole are required for an antigenic envelope to function as a protein protective envelope^16,27.

As oxygen carrier:

Khopade et al., prepared hydroxyapatite core, coated with trehalose followed by hemoglobin adsorption. Aquasomes have a good potential to be employed as an oxygen transporter and can continue their activity for 30 days, according to in vivo tests in rats. It was discovered that the oxygen-carrying capacity of the aquasome formulation was comparable to that of fresh blood. The timing of blood coagulation was not affected, and the aquasome formulations did not cause hemolysis of the red blood cells^7,28.

MARKETED PRODUCTS:

Aquasome-based delivery of FDA-approved recombinant proteins:

Table 3: FDA approved marketed products²⁹

Trade Name	Recombinant Product	Marketed Name	Manufacturer	Ref
Activase	Tissue plasminogen activator	Activase, Cathflo, Cathflo Activase	Boehringer Ingelheim	30
Cerezyme	Glucocerebrosidase	Cerezyme	Genzyme, Sanofi	30
Epogen/ procrit	Erythropoietin	Epoetin Alfa Hexal, Eporatio, Eprex	Amgen, Johnson and Johnson	30
Fabrazyme	Galactosidase A	Fabrazyme	Sanofi and Genzyme	30
Gonal-f	Follicle stimulating hormone	Bemfola, Follistim, Menopur, Ovaleap, Pergoveris, Puregon	Merck, EMd Serono	30
Herceptin	Anti-HER 2 humanized mAb	Herceptin Hylecta, Herzuma, Kanjinti, Ontruzant	Genentech, Amgen	30
Luveris	Luteinizing hormone	Luveris, Pergoveris	Merck Europe B.V, EMD Serono	30
Myozyme	Acid –glucosidase	Lumizyme, Myozyme	Genzyme, Sanofi	30
Novoseven	Clotting factorVII a	Niastase RT, Novoseven, Sevenfact	Novo Nordisk	30
Ovidrel	Human chronic gonadotropin	Ovitrelle, Pregnyl	Merck, EMD Serono	30
Raptiva	Anti-CD11a humanized mAb	Raptiva	Serono Europe Limited	30
Simulect	Anti-IL2receptor-chimeric mAb	Simulect	Novartis	30
Thyrogen	Thyrotropin	Thyrogen	Genzyme	30

MARKET POTENTIAL:

Aquasomes are self-assembled nanoparticle delivery systems with three layers that are intended to successfully transport bioactive substances while maintaining their structural integrity. Aquasomes has got a quite versatile application potential as a carrier for delivery of vaccines, haemoglobin, drugs, dyes and enzymes. Recent studies focused on the determination of the suitable aquasomal composition for protein and peptide drug delivery. One of the newest delivery systems for bioactive chemicals such peptides, proteins, hormones, antigens, and genes to particular areas is aquasomes.

FUTURE PROSPECTS:

Aquasomes can be considered as the most recently developed drug delivery system which holds a future potential in transdermal drug delivery. Aquasomes suspension contains a colloidal range of biodegradable nanoparticles hence, there is a high chance of accumulation in muscles and liver. The development of aquasomes and their characteristics allow for the efficient delivery of a wide range of therapeutic compounds.

CONFLICT OF INTEREST:

In relation to this inquiry, the authors have no conflicts of interest.

ACKNOWLEDGEMENT:

Authors are thankful to Bharati Vidyapeeth’s College of Pharmacy, Sector-8, C.B.D., Belapur, Navi Mumbai for providing the necessary facilities.

REFERENCES:

1. Banerjee S, Sen KK. Aquasomes: A novel nanoparticulate drug carrier. Journal of Drug Delivery Science and Technology. 2018; 43: 446-52. doi: 10.1016/j.jddst.2017.11.011

2. Kossovsky N, Gelman A, Sponsler EE, Hnatyszyn HJ, Rajguru S, Torres M et al.Surface-modified nanocrystalline ceramics for drug delivery applications. Biomaterials. 1994; 15(15): 1201–07. doi:10.1016/0142-9612(94)90270-4

3. Prasanthi NL, Manikiran SS, Krishna CS, Rao NR. Aquasomes: role to deliver bioactive substances. Research Journal of Pharmaceutical Dosage Forms and Technology. 2010; 2(6): 356-360

4. Pandey RS, Sahu S, Sudheesh MS, Madan J, Kumar M, Dixit VK. Carbohydrate modified ultrafine ceramic nanoparticles for allergen immunotherapy. International Immunopharmacology. 2011; 11(8): 925–31. doi: 10.1016/j.intimp.2011.02.004 .

5. Ilhan M, Gultekin HE, Rencber S, Senyigit Z, Aydin HH. Chapter 12 - Aquasomes: a novel platform for drug delivery. Systems of Nanovesicular Drug Delivery. Academic Press. 2022; 191-206. doi:10.1016/B978-0-323-91864-0.00020-6.

6. Gupta AK, Gupta D, Gupta V. Aquasomes: A Self-Assembled Nano-Particulate Carrier System. International Journal of Current Research and Review. 2021; 13(4): 44-52. doi: 10.31782/IJCRR.2021.13427.

7. Asfour MH. Advanced trends in protein and peptide drug delivery: a special emphasis on aquasomes and microneedles techniques. Drug Delivery and Translational Research. 2021; 11(1): 1-23. doi:10.1007/s13346-020-00746-z .

8. Umashankar MS, Sachdeva RK, Gulathi M. Aquasomes: a promising carrier for peptides and protein delivery. Nanomedicine: Nanotechnology, Biology, and Medicine. 2010; 6(3): 419–26. doi:10.1016/j.nano.2009.11.002 .

9. Jagdale S, Karekar S. Bird's eye view on aquasome: Formulation and application. Journal of Drug Delivery Science and Technology. 2020; 58: 1-14. doi: 10.1016/j.jddst.2020.101776 .

10. Yadav AR, Mohite SK. Aquasomes as a self assembling nanobiopharmaceutical carrier system for bio-active molecules. Research Journal of Topical and Cosmetic Sciences. 2020; 11(2): 89-94.

11. Mummidi V, Govada P, Mastanamma SK. Poly hydroxy oligomer coated dolutegravir aquasomes and method thereof. PCT/IN2021/050832; 2021.

12. Srinivasan M, Shankar U. Gel Formulating for treating diabetic foot ulcers infections. United States Patent, USS16/221,540; 2020.

13. Chaudhary JS. Aquasomes; a new approach for delivering therapeutics: an overview. Asian Journal of Pharmaceutics. 2018; 12(2): doi: 10.22377/ajp.v12i02.2369

14. Narang N. Aquasomes: Self-assembled systems for the delivery of bioactive molecules. Asian Journal of Pharmaceutics. 2012; 6(2): 95-100. doi:10.4103/0973-8398.102931

15. Khandbahale SV, Saudagar RB. Nanoparticle- a review . Asian Journal of Research in Pharmaceutical Sciences. 2017; 7(3):162-172.

16. Goyal AK, Khatri K, Mishra N, Mehta A, Vaidya B, Tiwari S, et al. Aquasomes—A Nanoparticulate Approach for the Delivery of Antigen. Drug Development and Industrial Pharmacy. 2008; 34(12): 1297-1305. doi: 10.1080/03639040802071661.

17. Kommineni S, Ahmad S, Vengala P, Subramanyam CVS.Drug Development and Industrial Pharmacy. 2012; 38(5): 577–86. doi:10.3109/03639045.2011.617884

18. Viswanathan S, Kumar NV, Srinivasan P, Prabhu S. Nanoparticle-mediated drug delivery systems. Research Journal of Engineering and Technology. 2013; 4(4): 295-299.

19. Kossovsky N. Biomolecular Delivery Using Coated Nanocrystalline Ceramics (Aquasomes), Chapter 23, ACS Publications, 1996; 622: 334-50.

20. Dange YD, Honmane SM, Patil PA, Gaikwad UT, Jadge DR. Nanotechnology, nanodevice drug delivery system: a review. Asian Journal of Pharmacy and Technology. 2017; 7(2): 63-71.

21. Sangar OS, Patil AC, Payghan SA. Nanoparticles: as a nano based drug delivery system. Asian Journal of Research in Pharmaceutical Sciences. 2022; 12(1):11-6.

22. Seenuvasan M, Balaji N, Kumar MA. Review on enzyme loaded magnetic nanoparticles. Asian Journal of Pharmacy and Technology. 2013; 3(4): 200-208.

23. Goyal AK, Rawat A, Mahor S, Gupta PN, Khatri K, Vyas SP. Nanodecoy system: a novel approach to design hepatitis B vaccine for immune potentiation. International Journal of Pharmaceutics. 2006; 309(1-2): 227–33. doi:10.1016/j.ijpharm.2005.11.037

24. Merlin NJ, Shaji S, Sukesh TN. Nano technology –potential applications in medicine. Asian Journal of Research in Pharmaceutical Sciences. 2011; 1(2): 31-35.

25. Sutariya V, Patel P. Aquasomes: a novel carrier for drug delivery. International Journal of Pharmaceutical Sciences and Research. 2012; 3(3): 688-94. doi:10.13040/IJPSR.0975-8232

26. Kulkarni S, Prabhakar B, Shende P. Aquasomes: Advanced vesicular-based nanocarrier systems. Current Pharmaceutical Design. 2022; 28(29): 2404-14. doi: 10.2174/1381612828666220728112741.

27. Kengar MD, Jadhav AA, Kumbhar SB, Jadhav RP. A review on nanoparticles and its application. Asian Journal of Pharmacy and Technology. 2019; 9(2): 115-124

28. Marnoor SA. A review on nanoparticles and its applications in therapeutics as a novel drug delivery system. Asian Journal of Pharmacy and Technology. 2019; 9(2): 125-128.

29. Hada NK, Ashawat MS. Aquasome: A Self-Assembling Supramolecular and Nanoparticulate Carrier System for Bio-Actives. Research Journal of Pharmaceutical Dosage Forms and Technology. 2013; 6(1): 50-3.

30. Drug Bank Online | Database for Drug and Drug Target Info. https://go.drugbank.com/drugs/ (accessed on 08/08/2022).

Received on 16.09.2022 Modified on 22.02.2023

Research J. Pharm. and Tech 2023; 16(12):6081-6086.

DOI: 10.52711/0974-360X.2023.00987