Enhancing biopharmaceutical attributes of phospholipid complex-loaded nanostructured lipidic carriers of mangiferin: Systematic development, characterization and evaluation

https://doi.org/10.1016/j.ijpharm.2016.12.044Get rights and content

Abstract

Mangiferin (Mgf), largely expressed out from the leaves and stem bark of Mango, is a potent antioxidant. However, its in vivo activity gets tremendously reduced owing to poor aqueous solubility and inconsistent gastrointestinal absorption, high hepatic first-pass metabolism and high P-gp efflux. The current research work, therefore, was undertaken to overcome the biopharmaceutical hiccups by developing the Mgf-phospholipid complex (PLCs) loaded in nanostructured lipidic carriers (NLCs). The PLCs and NLCs were prepared using refluxing, solvent evaporation and hot emulsification technique, respectively with three molar ratios of Mgf and Phospholipon 90 G, i.e., 1:1; 1:2; and 1:3. The complex was evaluated for various physicochemical parameters like drug content (96.57%), aqueous solubility (25-fold improved) and oil-water partition coefficient (10-fold enhanced). Diverse studies on the prepared complex using FTIR, DSC, PXRD and SEM studies ratified the formation of PLCs at 1:1 ratio. The PLCs were further incorporated onto NLCs, which were systematically optimized employing a face centered cubic design (FCCD), while evaluating for particle size, zeta potential, encapsulation efficiency and in vitro drug release as the CQAs. Caco-2 cell line studies indicated insignificant cytotoxicity, and P-gp efflux, while bi-directional permeability model and in situ perfusion studies specified enhanced intestinal permeation parameters. In vivo pharmacokinetic studies revealed notable increase in the values of Cmax (4.7-fold) and AUC (2.1-fold), respectively, from PLCs-loaded NLCs vis-à-vis Mgf solution. In a nutshell, the promising results observed from the present research work signify enhanced biopharmaceutical attributes of the novel PLCs-loaded NLCs for potentially augmenting the therapeutic efficacy of Mgf.

Introduction

Of late, mangiferin (Mgf), a polyphenolic C-glycoside has been gaining enormous recognition in pharmaceutical and food industries for its nutritional and therapeutic values. It possesses several pharmacological activities like antioxidant (Rodriguez et al., 2006), anti-inflammatory (Saha et al., 2016), anti-viral (Zheng and Lu, 1990), analgesic (Dar et al., 2005), immunomodulatory (Leiro et al., 2004, Sarkar et al., 2004), antidiabetic (Ojewole, 2005), besides other’s. Amongst all these, the ability of Mgf to act as a potent antioxidant by scavenging free radicals is known to be of immense importance for preventing the potential DNA damage (Zhang, 2013) and for inhibiting the growth of cancer cells by inducing cellular apoptosis (Minglin et al., 2016, Takeda et al., 2016).

Despite being a potent phytomolecule, the use of Mgf is restricted due to its low and variable oral bioavailability (about 1.5 to 5.0%), usually ascribed to its poor aqueous solubility (0.1 to 0.3 mg/mL), low lipophilicity, high P-gp efflux, considerable first-pass metabolism and cytochrome P-450 mediated metabolism in gut enterocytes (Basheer and Kerem, 2015, Khurana et al., 2016). Besides, Mgf display rapid clearance from the body via glucuronidation of the hydroxyl groups present at position-6 and -7 of its xanthone moiety, which, in turn, tends to reduce its potential antioxidant and anticancer activities, due to shorter half-life (van der Merwe et al., 2011).

Mgf, till date, is marketed in the form of conventional tablets, along with other antioxidants. However, such formulations exhibit limited fruition for potentiating the desired therapeutic efficacy of Mgf owing to the aforementioned biopharmaceutical hiccups. A score of formulation strategies including solid dispersions (Qin et al., 2014), cyclodextrin inclusion complex (da Rocha Ferreira et al., 2013, Yang et al., 2013), crystal engineering and salt formation (Teng and Wu, 2012), phospholipid complex (Ma et al., 2014) and spray-dried encapsulation (Souza et al., 2013) have already been reported in literature. These systems, however, have yielded limited improvement in the biopharmaceutical attributes of Mgf, plausibly owing to mere improvement in solubility and dissolution behavior. Keeping in view of the potential limitations of the existing formulation techniques, the development of newer and more effective formulation strategies for Mgf is highly called for.

The phospholipid systems, in this context, have lately been explored for enhancing the biopharmaceutical performance and therapeutic efficacy of bioactives with poor hydrophilicity and lipophilicity (Renukuntla et al., 2013). As phospholipids constitute a major part of the bio-membrane, these hold good biocompatibility and act as a carrier for delivering drugs across the biological barriers (Renukuntla et al., 2013). The amphiphilic nature of phospholipids tends to provide enhanced solubility and permeability of the drugs, thus improving their oral biopharmaceutical performance (Pichot et al., 2013). Other stellar merits of phospholipidic formulations include ease of preparation, along with high drug loading capacity and long-term stability. Several, literature reports suggest that incorporation of drug-phospholipid complexes into the nanostructured systems can further have remarkable increase in the biopharmaceutical performance of the drugs (Singh et al., 2012, Rawat et al., 2013, Xia et al., 2013).

The nanosystems composed of lipids not only enable the increased solubility and bioavailability of less soluble drugs, but also augment release and permeability of drugs, along with gastric protection against degradation from luminal enzymes (Renukuntla et al., 2013). Indeed, nanocarriers such as lipid micelles (Neuberg et al., 2015), solid lipid nanoparticles (Mukherjee et al., 2009, Bunjes, 2010), nanostructured lipidic carriers (NLCs) (Khan et al., 2015) and lipid nanocapsules (Sánchez-Moreno et al., 2012) have already been reported to hold immense promise for oral delivery of poorly water-soluble bioactives. Amongst these nanocarriers, the NLCs are considered to be of wider interest owing to their high drug-loading efficiency and drug stability (Naseri et al., 2015, Shah et al., 2016). Accordingly, the current work on the proposed novel technology was undertaken, wherein the PLCs of Mgf were incorporated into NLCs, for further enhancing the biopharmaceutical attributes. The nature of the intended research work on Mgf has not been reported in literature so far.

The present research work, therefore, involves the investigation of novel dual formulation strategy involving the use of PLCs-loaded NLCs for improving the oral bioavailability of Mgf. In this regard, PLCs were prepared and extensively characterized through FTIR, DSC, PXRD, SEM, and molecular docking studies for confirming the formation of complex. Further, intestinal absorption and permeability studies on the developed formulations have been performed on Caco-2 bidirectional assay model. Subsequently, intestinal perfusion studies and pharmacokinetic studies were also performed on Sprague-Dawley (SD) rats for evaluating the improvement in permeability, bioavailability and absorption parameters.

Section snippets

Materials and methods

Mgf was provided ex-gratis by International Association on Mangiferin Research (IAMR), Nanning, Guangxi, China. Phospholipon 90 G (PL) (soybean lecithin at 90% of phosphatidylcholine) was provided as gift sample by M/s Phospholipon GmbH, Germany. Labrafil M 2125 and Compritol 888 ATO were gifted by M/s Gattefosse, Cedex, France. All the solvents were procured from Merk Pvt. Ltd., India.

Results and discussion

The PLCs was formed employing stoichiometric ratios of 1:1, 1:2, and 1:3 and the following parameters were estimated to screen the best ratio.

Conclusions

A dual formulation strategy was adopted in order to enhance the biopharmaceutical attributes of a very potent antioxidant, Mgf. A minimum stiochiometric ratio of Mgf:PL (1:1) was observed to be good enough to form the complex on the basis of drug content, solubility and oil-water partition coefficient. Higher solubility and permeability values may lead to increased intestinal uptake and permeability, which is indeed a big limitation for oral absorption. Hence, the PLCs showed different

Declaration of interest

Authors declare no conflict(s) interest.

Acknowledgements

The financial grants received from the University Grant Commission (UGC), New Delhi, India, to Ms Rajneet Kaur Khurana under the RFMS scheme-F. No. 5-94/2007(BSR) is deeply acknowledged. Also, the services provided by SEM department of Central Instrumental Laboratory, Panjab University is highly appreciable. Use of Biomedical research facilities at University of Central Lancashire is acknowledged. We are also thankful for the generous supply of hard gelatin capsules shells ex-gratis by M/s ACG

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