FORMULATION, OPTIMIZATION AND IN VITRO EVALUATION OF 5-FLUOROURACIL LOADED LIQUORICE CRUDE PROTEIN NANOPARTICLES FOR SUSTAINED DRUG DELIVERY USING BOX-BEHNKEN DESIGN
Keywords:Nanoparticle, Plant protein, Desolvation method, Particle size, Zeta potential, Entrapment efficiency, Drug release
Objective: To formulate, optimize and evaluate 5-fluorouracil loaded liquorice crude protein nanoparticles for sustained drug delivery using Box-Behnken design.
Methods: 5-fluorouracil (5-FU) loaded liquorice crude protein (LCP) nanoparticles were prepared by desolvation method using ethanol-water (1:2 ratio), Tween-80 (2%v/v) as stabilizing agent and gluteraldehyde (8% v/v) as cross linking agent. The optimization of prepared nanoparticles was carried out using Box-Behnken design with 3 factors 2 levels and 3 responses. The independent variables were A)5-FU concentration B)LCP concentration and C) sonication time while the responses were R1) Drug entrapment efficiency R2) Drug loading efficiency and R3) Particle size. The correlation between factors and responses were studied through response surface plots and mathematical equations. The nanoparticles were evaluated for FTIR, physicochemical properties like particle size and zeta potential by Photon correlation spectroscopy (PCS) and surface morphology by TEM. The entrapment efficiency, drug loading efficiency and in vitro drug release studies in PBS pH 7.4 (24 h) were carried out. The observed values were found to be in close agreement with the predicted value obtained from the optimization process.
Results: 5-fluorouracil loaded LCP nanoparticles were prepared by desolvation method, the optimization was carried out by Box-Behnken design and the final formulation was evaluated for particle size (301.1 nm), zeta-potential (-25.8mV), PDI(0.226), with entrapment efficiency (64.07%), drug loading efficiency (28.54%), in vitro drug release (65.2% in 24 h) respectively. The formulated nanoparticles show Higuchi model drug release kinetics with sustained drug delivery for 24 h in pH7.4 buffer.
Conclusion: The results were proved to be the most valuable for the sustained delivery of 5-Fluorouracil using liquorice crude protein as carrier. 5-FU–LCP nanoparticles were prepared using Tween-80 as stabilizing agent and gluteraldehyde as cross-linking agent to possess ideal sustained drug release characteristics.
Lakshmana Rao Atmakuri, Suneetha Dathi. Current trends in herbal medicines. J Pharm Res 2010;3:109-13.
Biswajit Mukherjee, Kousik Santra, Gurudutta Pattnaik, Soma Ghosh. Preparation, characterization and in vitro evaluation of sustained release protein loaded nanoparticles based on biodegradable polymers. Int J Nanomed 2008;3:487-96.
V Kusum Devi, Nimisha Jain, Kusum S Valli. Importance of novel drug delivery system in herbal medicine. Pharmacogn Rev 2010;4:27–31.
Lekha Nair K, Sankar Jagadeeshan, S Asha Nair, GS Vinod Kumar. Biological evaluation of 5FU nanoparticles for cancer chemotherapy and its dependence on the carrier, PLGA. Int J Nanomed 2011;6:1685-97.
Amir Maghsoudi, Seyed Abbas Shojaosadati, Ebrahim Vasheghani Farahani. 5-Fluorouracil-loaded BSA nanoparticles: formulation optimization and in vitro release study. AAPS PharmSciTech 2008;9:1092-6.
Amrit Rai, Josephine Jenifer, Ravi T Prakash. Nanoparticles in therapeutic applications and role of albumin and casein nanoparticles in cancer therapy. Asian Biomed 2017;11:3-20.
Ahmed E, Wael Mohammed Samy, Nazik A. Albumin based nanoparticles as potential controlled release drug deliver systems. J Controlled Release 2011;157:168-82.
Ann M Bode, Zigang Dong. Chemopreventive effects of liquorice and its components. Curr Pharmacol Reports 2015;1:60-71.
Shibata S. A drug over the millennia: pharmacognosy chemistry and pharmocology of liquorice. Yakugaku Zasshi 2000;120:849-62.
Hemraj Vashist, Diksha Sharma. Pharmacognostical aspects of glycerrhiza glabra. Asian J Pharm Clin Res 2013;6:55-9.
Xiao Ting Chu, Joseph de la Cruz, Seong Gu Hwang, Heeok Hong. Tumorigenic effects of endocrine-disrupting chemicals are alleviated by licorice (Glycyrrhiza glabra) root extract through suppression of AhR expression in mammalian cells. Asian Pac J Cancer Prev 2014;15:4809-13.
Mukne Alka Pravin, Viswanathan Vivek, Pharande Rajesh Raghunath, Bannalikar Anilkumar Sadashivrao. Acute toxicity studies of nano-formulations of glycyrrhiza glabra extract in swiss albino mice. World J Pharm Pharm Sci 2017;6:820-9.
Swati Chauhan, Neha Gulati, Upendra Nagaich. Glycyrrhizic acid, extraction, screening and evaluation of an anti-inflammatory property. Ars Pharm 2018;59:61-7.
Geetha VS, Malarkodi Velraj. Development and optimization of liquorice crude protein nanoparticles from glycyrrhiza glabra L. J Innovations Appl Pharm Sci 2020;5:1-9.
Sherif EA Badr, Dina M Sakr, Sanaa A Mahfouz, Mohamed S Abdelfattah. Licorice (Glycyrrhiza glabra L.): chemical composition and biological impacts. RJPBCS 2013;4:606-21.
Rajinikanth S, Kumarappan Chidambaram. Topical nano-delivery of 5-fluorouracil: preparation and characterization of water-in-oil nanoemulsions. Trop J Pharm Res 2016;15:2311-9.
Li Sun, Yunna Chen, Yali Zhou, Dongdong Guo, Yufan Fan, Fangyan Guo, et al. Preparation of 5 fluorouracil loaded chitosan nanoparticles and study the sustained release in vitro and in vivo. Asian J Pharm Sci 2017;17:1-21.
Shashank Tummala M, N Satish Kumar, Ashwati Prakasha. Formulation and characterization of 5-fluorouracil enteric coated nanoparticles for sustained and localized release in treating colorectal cancer. Saudi Pharm J 2015;23:308-14.
Vijayan Venugopal, K Jayaraja Kumar, S Muralidharan, S Parasuraman, P Vasanth Raj, K Venkates Kumar. Optimization and in vivo evaluation of isradipine nanoparticles using box-behnken design surface response methodology. Open Nano 2016;1:1–15.
Vijay Bahadur Kumal, Chhitij Thapa, Prakash Ghimire, Pradyumna Chaudhari, Jitendra Yadhav. Formulation and optimization of enalapril maleate loaded floating microsphere using box–behnken design: in vitro study. J Appl Pharm Sci 2020;10:95–104.
Kousalya Prabahar, Ubaidulla Udhumansha, Mona Qushawy. Optimization of thiolated chitosan nanoparticles for the enhancement of in vivo hypoglycemic efficacy of sitagliptin in streptozotocin-induced diabetic rats. Pharmaceutics 2020;12:300.
Sailaja Pb, Jeevana Jyothi B. Development and in vitro evaluation of 5-fluorouracil nanoparticles by salting out technique. Asian J Pharm Clin Res 2020;13:166-70.
T Mallamma, Thippeswamy BS, Bharathi DR, Snehalatha, Nagaraja TS. Formulation and evaluation of 5-flurouracil loaded hsa nanoparticle for controlled drug delivery. Int J Adv Res 2013;7:23-30.
Chinmay G Hiremath, Mahadevappa Y Kariduraganavar, Murigendra B Hiremath. Synergistic delivery of 5‑fuorouracil and curcumin using human serum albumin‑coated iron oxide nanoparticles by folic acid targeting. Prog Biomater 2018;7:297–306.
Abdullah Alomrani, Mohamed Badran, Gamaleldin I Harisa, Mohamed ALshehry, Moayed Alhariri, Aws Alshamsan, et al. The use of chitosan-coated flexible liposomes as a remarkable carrier to enhance the antitumor efficacy of 5-fluorouracil against colorectal cancer. Saudi Pharm J 2019;27:603-11.
Ghasemi Z, Dinarvand R, Mottaghitalab F, Esfandyari Manesh M, Sayari E, Fatemeh atyabi aptamer decorated hyaluronan/chitosan nanoparticles for targeted delivery of 5-fluorouracil to MUC1 overexpressing adenocarcinomas. Carbohydrate Polymer 2014;121:190-8.
Nipaporn Ngernyuang, Wunchana Seubwa, Sakda Daduang, Patcharee Boonsiri, Temduang Limpaiboon, Jureerut Daduang. Targeted delivery of 5-fluorouracil to cholangiocarcinoma cells using folic acid as targeting agent. Mater Sci Eng C Mater Biol Appl 2016;60:411-5.
Shushil Bhusal, Khaga Raj Sharma. Evaluation of the antioxidant and antibacterial activity of glycyrrhiza glabra root extracts. Asian J Pharm Clin Res 2020;13:166-70.
Indian Pharmacopoeia 2018. 8th edition. Government of India, Ministry of Health and Family Welfare. Indian Pharmacopoeial Commission, Ghaziabad; 2018. p. 1969-71.
Li Jing Ke, Guan Zhen Gao, Yong Shen, Jian Wu Zhou, Ping Fan Rao. Encapsulation of aconitine in self-assembled licorice protein nanoparticles reduces the toxicity in vivo. Nanoscale Res Lett 2015;10:449.
Vyas SP, Khar RK. Targeted and controlled drug delivery: novel carrier systems. CBS Publications 2004;1:383-410.
Becket AH, Stenlake JB. Practical pharmaceutical chemistry. Part–II. 4th edition. CBS Publishers and Distributors, New Delhi; 2007. p. 965-98.
Dr Pulok, K Mukherjee. Quality control of herbal drugs, Horizon Pharmaceutical Publishers; 2012.
KD Tripathi. Essentials of medical pharmacology. 8th Edition. Jaybee brothers medical publishers; 2018.