Formulation and Characterization of phytophospholipid complex of Andrographis paniculata extract

Abstract

Active constituents extracted from various parts of the plants have wide therapeutic potential for the treatment of diseases. Number of these compounds obtained elicits positive in vitro pharmacological effect; however poor in vivo absorption of polyphenolic compounds limited the widespread applicability. One of the novel strategies to improve the bioavailability and thereby oral absorption of these compounds is the use of phytophospholipid complexes. These complexes are formed by combining phyto constituent in definite molar ratio with phospholipids under certain specific conditions. This approach has overcome the obstacle of poor oral bioavailability of numbers of active constituents most of which are polyphenolic compounds (1). Some of the biologically active polyphenolic constituents of plants show affinity for the aqueous phase and cannot pass through biological membranes, such as hesperidin. By contrast, others have high lipophilic properties and cannot dissolve in aqueous gastrointestinal fluids, such as curcumin and rutin. Phyto-phospholipid complexes can not only improve the solubility of lipophilic polyphenols in aqueous phase but also improves the membrane penetrability of hydrophilic compounds from aqueous phase. Furthermore, the production of complexes can protect polyphenols from destruction by external forces, such as hydrolysis, photolysis, and oxidation therefore this novel technique has received much attention in recent years (2). Nandhani S (2021) developed and optimized Vasaka-loaded phytosomes by thin-layer hydration technique using systematic quality by design approach. Box-Behnken design (3² factorial designs) using Design-Expert software was employed to optimize phytosome wherein phosphatidylcholine concentration (X₁), stirring temperature (X₂), and stirring time (X₃) were selected as independent variables. Yield (%), particle size (nm), and entrapment efficiency (%) were evaluated as responses. The optimized phytosome was characterized by studying the surface morphology such as FE-SEM and TEM analysis, thermal characteristics by thermal gravimetric analysis and spectral and diffraction studies by FTIR and XRD analysis and studying the dissolution behaviour of phytosome by in vitro release study(3). Cheng Chi et al (2020) developed a phytosome-nanosuspensions for silybin-phospholipid complex loading, termed SPCs-NPs, for hepatoprotection efficacy. This formulation has shown an increased dissolution rate and more potent hepatoprotective effects in pharmacodynamic assessments (4). David Pastorelli et al (2018) developed Phytosome complex of curcumin as complementary therapy of advanced pancreatic cancer (PC). Experimental data suggested that curcumin in complexed form with phospholipids can be used in the treatment of PC as a complementary therapy to Gemcitabin (5). Andrographis paniculata (A. paniculata) is a medicinal plant found in southern part of India traditionally used as anti-inflammatory and anti-bacterial herb. It exhibits diverse pharmacological activities, including anti-inflammation, anti-cancer, anti-obesity, anti-diabetes, Hepatoprotective and other activities. Oral administration of A. paniculata (100-200mg/kg) showed significant dose dependent hepatoprotective effect assessed in terms of biochemical and histopathological parameters.  The principal constituent of Andrographis paniculata is andrographolid it is a diterpenoid lacton present in leaves, poor bioavailability, short biological half life and inappropriate tissue distribution limited its therapeutic applications (6). The present work is aimed to formulate and characterize phytophospholipid complex of methanolic extract of leaves of Andrographis paniculata by anti-solvent precipitation technique. Andrographis paniculata Burm. F. Nees aerial parts (stems and leaves) of phytosomes were prepared by using anti solvent evaporation method. Andrographis powder was placed in a round bottom flask. The phytosomal formulations were prepared (F1-F4) in different molar ratios of (plant extract: soy lecithin) 1: 0.5, 1:1, 1:1.5, 1:2. Exact quantity of plant extract and soy lecithin was kept in 100 ml round bottom flask and then refluxed with 20 ml of dichloromethane at 60˚C for 2hours. The mixture was concentrated to 5 ml. Hexane was added carefully to get mixture precipitated which was filtered, collected and stored in a vaccum dessicator. The dried precipitate was crushed in a pestle mortar. The prepared phytosomes were than characterised. The characterization of prepared phytosomes was done by using particle size analysis PDI (Polydispersity index), DSC (Differential Scanning Calorimetry), XRD (X-Ray Diffraction), SEM (Scanning electron microscope) method, Entrapment efficiency (EE) and in-vitro drug release studies were done to evaluate the parameters of phytosome complexes. The formulation F4 possesses good particle size with PDI 0.567 and zeta potential was in range of -31.57±0.2. This indicates that phytosomal formulation possess good stability. In vitro drug released studies carried out for the period of 6 hrs indicated that the formulation F4 gives 88% drug release which was found to be sustained. Surface morphology studies indicated drug extract particles associated with phospholipids were uniform and spherical in shape. Phytosomes of Andrographis paniculata extract have got ability, that can be used to enhances solubility and stability studies and hence oral bioavailability for which elaborative in vivo studies need to be carried out so that a novel formulation having improved pharmacological activity can be developed.