Combination approach of nanocrystals and dissolving microneedles for improved pharmacokinetic profiles of albendazole in treatment of cyctic echinococcosis

Andi Dian Permana¹, Alejandro J. Paredes², Fabiana Volpe-Zanutto^2,3, Muh. Nur Amir⁴, Ismail Ismail⁵, Muh. Akbar Bahar⁴, Sumarheni⁶, Santiago Daniel Palma⁷, Ryan F. Donnelly^2*

1. Department of Pharmaceutics, Faculty of Pharmacy, Hasanuddin University, Makassar 90245, Indonesia
2. School of Pharmacy, Queen's University Belfast, Medical Biology Centre, 97 Lisburn Road, Belfast.  BT9 7BL, UK
3. Faculty of Pharmaceutical Sciences, R. Cândido Portinari, 200 - Cidade Universitária, Campinas - SP, 13083-871, University of Campinas, Brazil
4. Department of Pharmacology and Toxicology, Faculty of Pharmacy, Universitas Hasanuddin, Makassar 90245
5. Department of Phytochemistry, Faculty of Pharmacy, Universitas Hasanuddin, Makassar 90245, Indonesia
6. Department of Clinical Pharmacy, Faculty of Pharmacy, Universitas Hasanuddin, Makassar 90245, Indonesia
7. Unidad de Investigación y Desarrollo en Tecnología Farmacéutica (UNITEFA), CONICET and Departamento de Ciencias Farmacéuticas, Facultad de Ciencias Químicas, Universidad Nacional de Córdoba, Argentina. Haya de la Torre y Medina Allende, X5000XHUA, Córdoba, Argentina.

Introduction

Cystic echinococcosis (CE) is a devastating zoonosis which affects both humans and animals. CE is caused by Echinococcus spp., commonly Echinococcus granulosus.^1,2 Presently, there are several choices for the treatment of CE. These include antiparasitic administration, surgical intervention, and percutaneous drainage therapy. The selection of treatments is dependent on the nature of the cysts in each patient.³ The administration of antiparasitic agents is the first treatment option where doctors are not available. Additionally, this is the only alternative in numerous inoperable circumstances, including cysts in the brain and cysts in patients who are immune-supressed.⁴  Currently, the benzimidazole carbamate derivate, albendazole (ABZ), is the most effective drug commonly used for the treatment of CE. However, ABZ has poor aqueous solubility, resulting in low bioavailability when administered orally. In addition, due to its low solubility in aqueous environments, the route of administration options are limited to the oral route.^5,6 Therefore, it is crucial to develop a formulation which can overcome the poor solubility.

When administered orally, ABZ encounters rapid first-pass metabolism in the liver, being transformed into ABZ-sulfoxide (ABZ-SX).⁷ Although ABZ and ABZ-SX show antiparasitic activities, ABZ has been reported to possess stronger affinity to parasite tubulins when compared to ABZ-SX, meaning that ABZ has higher activity than ABZ-SX.⁸ It has also been reported that the viability of cysts in NMRI mice infected by Echinococcus granulosus was lower after the administration of ABZ compared to those with the administration of ABZ-SX.⁹ Accordingly, a new delivery approach which can avoid the rapid metabolism of ABZ in the liver may enhance the efficacy of ABZ in the treatment of CE. 

Transdermal delivery systems are one of the most favorable methods to improve the delivery of numerous drugs and are able to avoid hepatic first-pass metabolism,^7,10 Amongst various delivery approaches, dissolving microneedles (DMNs) have shown numerous advantages compared to other strategies,¹¹ since they by-pass the skin’s stratum corneum.¹² In the presented study, we develop, for the first time, the combinatorial approach of nanocrystals and DMNs for the intradermal administration of ABZ using to improve its bioavailability.

Experimental methods:

Initially, the NCs were developed using top-down method using an ultra-small-scale media mill (Figure 1). Several characterizations were further performed, including particle size, polydispersity index, physical characteristics and drug release kinetics were further characterized. Subsequently, the NCs were loaded into DMNs (Figure 2) and characterizations of mechanical properties were carried out. Moreover, ex vivo dermatokinetic studies were also performed to examine the intradermal delivery of this approach. Finally, in vivo pharmacokinetic studies of ABZ in NCs after DMN administration in rats were compared to the conventional administration of ABZ in CE therapy.

Figure 1.  Schematic representation of NC preparation

Figure 2.  Schematic representation of two-layered DMN preparation

Results

Following several screenings, Pluronic^® F127 was selected as a suitable stabilizer, producing NCs with around 400 nm in size with narrow particle distribution. The crystallinity of ABZ was maintained as observed by DSC and XRD analysis. The NC approach was able to improve the dissolution percentage of ABZ by approximately 3-fold. Furthermore, the incorporation of NCs into DMNs using the combination of poly(vinylpyrrolidone) and poly (vinyl alcohol) formed sharp needles with sufficient mechanical strength and insertion properties. Dermatokinetic studies revealed that >25% of ABZ was localized in the dermis of excised neonatal porcine skin up to 48 h after DMN administration. In in vivo pharmacokinetic studies, the AUC and relative bioavailability values of ABZ delivered by NC-loaded DMNs were found to be significantly higher than those obtained after oral administration of coarse suspension of ABZ or ABZ-NCs, as well as DMNs delivering coarse ABZ as indicated by the relative bioavailability values of > 100% (Figure 3). Therefore, the combination approach developed in this study could maintain the systemic circulation of ABZ which could be possibly caused by avoiding the first-pass metabolism in the liver. This could be beneficial to improve the efficacy of ABZ in CE treatment.

According to the results discussed here, the incorporation of the NCs into DMNs was able to improve the bioavailability of ABZ, while avoiding the liver metabolism, as compared to the oral administration. Being delivered by DMNs, this technology is a safe handling, self-administered, painless and the most suitable options for CE treatment, especially the situation where it is extremely challenging to find a healthcare professional to treat this specific disease. Therefore, it is hypothesized that this novel approach could be beneficial as an alternative treatment of CE. The preclinical efficacy of ABZ-NCs in animal model of CE and NCs was evaluated previously using an oral treatment¹³, attaining an enhanced but limited therapeutic response against the parasite in chemoprophylaxis (which represents the rupture of the cysts during an extraction surgery) and post-infection (once the infection develops and the treatment is given) experiments.¹³ Solid dispersions of ABZ were also analyzed in the same model¹⁴ and lipid nanoparticles¹⁵. However, all the formulations showed limited efficacy in the CE model. In our study, as discussed previously, we were able to maintain the parent molecule in the circulation for the first time to the best of our knowledge. Therefore, we hypothesize that our findings represent a promising alternative to the current therapy of CE. However, further experiments should be considered, including toxicity studies, biocompatibility studies and an in vivo pharmacodynamic study to assess the effectiveness of this novel approach in the treatment of CE.

Figure 3. Mean plasma concentrations and time profiles of ABZ, ABZ-SX and ABZ-SN after oral administration of coarse suspension of ABZ (A) and ABZ-NCs (B), as well as after intradermal administration of coarse ABZ (C) and ABZ-NCs (D) using DMNs. The relative bioavailability values of ABZ delivered by DMN-NC compared to oral administration of coarse ABZ, oral administration of ABZ-NCs and DMN administration of ABZ-NCs (E) (means ± SD, n = 3).

Conclusions

This extensive work has shown the effectiveness of the combination of NCs and DMNs to overcome the issues of ABZ to potentially improve the treatment of CE. The formulation of ABZ into NCs, stabilized with Pluronic^® F127, enhanced the dissolution rate of ABZ, while maintaining the crystallinity of the drug.  Moreover, the formulation of ABZ-NCs into DMNs with adequate mechanical properties and skin insertion abilities resulted in enhancement of the concentration of ABZ retained in the dermis layer of the skin. Lastly, plasma pharmacokinetic assessment demonstrated that the intradermal delivery of ABZ-NCs delivered by DMNs could increase the relative bioavailability of the parent drug ABZ, and decrease the concentration of its metabolites, namely ABZ-SX and ABZ-SN, in comparison with the administration of coarse ABZ and ABZ-NCs via oral route, as well as DMN dosing of coarse ABZ. Therefore, this innovative approach could potentially lead to improvement of CE treatment. To prove its efficacy in the CE treatment, in vivo pharmacodynamic study will now be our next step.

Acknowledgements

This work was supported in part by Wellcome Trust grant number WT094085MA.

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