Development and Evaluation of Curcumin Peptide Concoction for Improving Brain Delivery and In-Vivo Efiicacy in Dementia Model

Abstract

Aim and Objective: The aim of study was to develop a curcumin peptide concoction to improve brain delivery along with solubility. The optimized concoctions are tested for in-vivo efficacy in dementia model. Introduction: The Indian Ayurvedic literature has shown a great potential in treating various disorders including life-threatening ones. Curcumin a novel compound present in rhizomes of Curcuma longa L. (Zingiberaceae; turmeric). Curcumin is oil soluble, practically insoluble in water at acidic and neutral pH while partial soluble in alkaline medium. Curcumin has been clinically studied till Phase-II but its water solubility and bioavailability are major apprehensions. In order to overcome curcumin solubility and bioavailability hitches different vectors, peptides, surface modifiers, size reduction techniques and other methods are commonly studied. Glutathione itself is a self-oxidizing agent thus lead to be a potential antioxidant molecule. While the other peptide was casein a phosphoprotein having a tertiary structure majorly found in milk and related products. Neurodegeneration is characterized by loss of neurons or neuronal structure leading to cell death. The main causes of neurodegeneration are mitochondrial deoxyribonucleic acid mutation, oxidative stress caused by advancing age, and inflammation. Alzheimer’s a type of pre-senile and senile dementia with 46.8 million people suffering worldwide in year 2015. In the current research, a surface modified curcumin was prepared using glutathione and casein as a peptide to increase its water solubility and bioavailability. Moreover, the concoction was further studied on in-vivo dementia model by use of aluminum chloride (AlCl3) as a chemical for induction. The prepared hypothesis is novel and the approach of use of this peptide is explored first time in improving aqueous solubility and bioavailability of curcumin in the management of dementia-related neurological disorder. Material and Methods: Curcumin (95%) was used as drug while casein (99%) and glutathione (99%) was used as peptide. Two type of formulation were prepared first by physical mixture and second by solvent evaporation technique using 9:1, 8:2 and 7:3 ratio of drug to peptide respectively in each. Moreover, a marketed formulation was selected based on its claimed aqueous solubility and brain bioavailability and kept as marketed standard for comparison in different estimations. The prepared concoctions and marketed formulations were evaluated via antioxidant analysis and some of them were estimated for in-vitro release profile to understand pH dependent variation of drug in terms to oral absorption while MTT assay was studied to estimate the viability of concoction on SHSY5Y cell line. Further, in-vivo pharmacokinetic study was estimated by giving single dose of prepared curcumin concoctions (2 g/kg equivalent) via oral administration that helps in knowing the bioavailability of curcumin at different tissues. Also, an in-vivo aluminum chloride based dementia model was studied to evaluate the effect of prepared concoctions in respect to behavioral, biochemical and histopathological changes.Result and Discussion: The concoction of curcumin-casein, curcumin-glutathione and curcumin-casein-glutathione had shown increase in water solubility in comparison to plain curcumin. The curcumin concoctions, observed that 9:1 is the ideal ratio using casein and glutathione as peptide using solvent evaporation technique. The concoction observed increase in solubility of 1432 times, 854 times, 868 times, and 326 times for curcumin-casein, curcumin-glutathione, curcumin-casein-glutathione complex, and marketed formulation respectively. Moreover, antioxidant parameter observed %inhibition of 68.46, 54.81, 64.85, 66.07, 73.25, and 65.96 respectively for DPPH assay. The pharmacokinetic estimation was measured by LC-MS/MS analysis in different tissues like brain, liver, lung, kidney, spleen were isolated at 20, 30, 40, 60,120 min time post dosage (2g/kg). The results showed curcuminoids in brain at 1.23, 1.19, 5.56, 74.41, and 1.38 ng/ml for curcumin, curcumin-glutathione, curcumin-casein, curcumin-casein-glutathione complex and marketed formulation respectively. The in-vivo Alzheimer’s study observed no change in body weight while Y-maze behavioral assessment test observed that in disease condition number of arm rotation was less while in normal condition arm rotation was more. Moreover, morris water maze behavioral assessment test observed that escape latency time (s) was less in normal control group while in diseased condition time required was more. The biochemical estimation like lipid peroxidation, glutathione, and total protein observed significant (p<0.01) change in all groups in comparison to disease control group. The total protein was estimated to measure the loss of blood either due to damage in systemic circulation or a disorder. The histopathology results observed change in hippocampus region by damaged cells and filaments of Aβ in the diseased control group in comparison to normal control group. Conclusion: The current research, concludes that curcumin prepared with casein and glutathione had found to increase its water solubility and also its bioavailability. The prepared concoctions observed as potential formulation in prevention or treatment in Alzheimer’s model. Thus, it needs to study further for different neurodegenerative disorders and models with pre-clinical and clinical efficacy. This research may help in exploring the molecule curcumin in a new way.