DESIGN, DEVELOPMENT AND INVITRO EVALUATION OF ERLOTINIB LOADED LIQUORICE CRUDE PROTEIN NANOPARTICLES BY BOX BEHNKEN DESIGN
Objective: To formulate and evaluate Erlotinib loaded Liquorice crude protein (LCP) nanoparticles from the powdered liquorice root (Glycyrrhiza glabra) using Box-Behnken design.
Methods: Erlotinib loaded liquorice crude protein nanoparticles were prepared by desolvation method using ethanol-water (1:2 ratio), Tween-20 (2%v/v) and gluteraldehyde (8% v/v) as cross linking agent. Box-Behnken design with 3 factors, 2 levels and 3 responses was used to optimize the prepared nanoparticles. The independent variables were taken as A) Erlotinib concentration B) LCP concentration and C) Incubation time with responses R1) Drug entrapment efficiency R2) Drug Release 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, particle size and zeta potential by Photon correlation spectroscopy (PCS) and surface morphology by TEM. The entrapment efficiency (%), and in vitro drug release(%) studies in PBS pH 7.4 (26 h) were carried out. The experimental values were found to be in close resemblance with the predicted value obtained from the optimization process. The invitro cytotoxicity studies of the prepared nanoparticles in lung cancer cell line (A 549) were studied with different concentrations at different time intervals.
Results: The average particle size, zeta potential, Polydispersity index (PDI) were found to be 292.1nm, -25.8 mV and 0.384 respectively. TEM image showed that the nanoparticles dispersed well with a uniform shape and showed not much change during storage. The invitro drug release showed 41.23% for 26 hrs in PBS (7.4) and release kinetics showed highest R2 value (0.982) for Korsmeyer-Peppas model, followed by 0.977 for Higuchi model. The invitro cytotoxicity of prepared nanoparticles in A 549 cell line showed good results with different concentrations at different time intervals.
Conclusion: Erlotinib (Erlo) is a BCS class II drug with poor solubility, poor bioavailability and selective tyrosine kinase inhibitor for non small-cell lung cancer (NSCLC) through oral administration. To improve the oral bioavailability and absorption of molecules, plant protein as carriers is used for developing drug delivery systems due to their proven safety. The optimization variables were Conc of Erlo, Conc. of LCP and Incubation time to get responses as drug entrapment efficiency, drug release and particle size. The compatibility between drug and LCP were evaluated by FTIR.
2. Dora CP, Trotta F, Kushwah V, Devasari N, Singh C, Suresh S, Jain S. Potential of erlotinib cyclodextrin nanosponge complex to enhance solubility, dissolution rate, in vitro cytotoxicity and oral bioavailability. Carbohydr Polym 2016;137:339-49.
3. Bhambere D, Shirivastava B, Sharma P, Gide P. Effect of Polymer and Formulation Variables on Properties of Self-Assembled Polymeric Micellar Nanoparticles. J Nanomedicine Biotherapeutic Discov 2014; 4:129.
4. Bhatia Saurabh. Nanoparticles Types, Classification, Characterization, Fabrication Methods And Drug Delivery Applications. Natural Polymer Drug Delivery Systems 2016; 33-93.
5. Shen Y, Li W. HA/HSA co-modified erlotinib-albumin nanoparticles for lung cancer treatment. Drug Des Devel Ther 2018;12: 2285-92.
6. Kim ST, Lee J, Kim JH, Won YW, Sun JM, Yun J, Park YH, Ahn JS, Park K, Ahn MJ. Comparison of gefitinib versus erlotinib in patients with nonsmall cell lung cancer who failed previous chemotherapy. Cancer 2010; 116:3025-33.
7. Sechler M, Cizmic AD, Avasarala S, Van Scoyk M, Brzezinski C, Kelley N, Bikkavilli RK, Winn RA. Non-small-cell lung cancer: molecular targeted therapy and personalized medicine - drug resistance, mechanisms, and strategies. Pharmgenomics Pers Med 2013; 6:25–36.
8. Çoban, Ö., De?im, Z. Development of Nanocochleates Containing Erlotinib HCl and Dexketoprofen Trometamol and Evaluation of In Vitro Characteristic Properties. Turkish journal of pharmaceutical sciences 2018;15:16–21.
9. Zhou X, Tao H, Shi KH. Development of a nanoliposomal formulation of erlotinib for lung cancer and in vitro/in vivo antitumoral evaluation. Drug Des Devel Ther 2017; 12:1-8.
10. Truong DH, Tran TH, Ramasamy T, Choi JY, Lee HH, Moon C, Choi HG, Yong CS, Kim JO. Development of solid self-emulsifying formulation for improving the oral bioavailability of erlotinib. Aaps Pharmscitech 2016; 17:466-73.
11. Mandal B, Mittal NK, Balabathula P, Thoma LA, Wood GC. Development and in vitro evaluation of core-shell type lipid-polymer hybrid nanoparticles for the delivery of erlotinib in non-small cell lung cancer. Eur J Pharm Sci 2016; 81:162-71.
12. Ajazuddin, Saraf S. Applications of novel drug delivery system for herbal formulations. Fitoterapia 2010; 81:680-9.
13. Thakur D, Abhilasha, Jain A, Ghoshal G. Evaluation of Phytochemical, Antioxidant and Antimicrobial Properties of Glycyrrhizin Extracted from Roots of Glycyrrhiza Glabra. Journal of Scientific and Industrial Research 2016; 75:487-94.
14. Shi L, Tang C, Yin C. Glycyrrhizin-modified O-carboxymethyl chitosan nanoparticles as drug vehicles targeting hepatocellular carcinoma. Biomaterials 2012; 33:7594-604.
15. Ke, L. J., Gao, G. Z., Shen, Y., Zhou, J. W., & Rao, P. F. Encapsulation of Aconitine in Self-Assembled Licorice Protein Nanoparticles Reduces the Toxicity In Vivo. Nanoscale research letters 2015;10: 449.
16. Kim B, Seo B, Park S, Lee C, Kim JO, Oh KT, Lee ES, Choi HG, Youn YS. Albumin nanoparticles with synergistic antitumor efficacy against metastatic lung cancers. Colloids Surf B Biointerfaces 2017;158:157-66.
17. 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.
18. Saravanan VS, Rao BM. Analytical Method Development and Validation for the Determination of Erlotinib Hydrochloride Bulk and in Pharmaceutical Dosage Form. Journal of Drug Delivery and Therapeutics 2013; 3:703-9.
19. M. Mathrusri Annapurna, B. Venkatesh and R. Krishna Chaitanya. Analytical Techniques for the Determination of Erlotinib HCl in Pharmaceutical Dosage Forms by Spectrophotometry. Chem Sci Trans 2014; 3:840-6.
20. Jahanban-Esfahlan A, Dastmalchi S, Davaran S. A simple improved desolvation method for the rapid preparation of albumin nanoparticles. Int J Biol Macromol 2021; 91:703-9.
21. Sravya Nella, Kiran Babu Uppuluri. Formulation and In-Vitro Evaluation of Piroxicam Loaded BSA Nanospheres by Desolvation, J Nanomed Nanotechnol 2015;6.
22. Yang KM, Shin IC, Park JW, Kim KS, Kim DK, Park K, Kim K. Nanoparticulation improves bioavailability of Erlotinib. Drug Dev Ind Pharm 2017; 43:1557–65.
23. Vrignaud S, Hureaux J, Wack S, Benoit JP, Saulnier P. Design, optimization and in vitro evaluation of reverse micelle-loaded lipid nanocarriers containing erlotinib hydrochloride. Int J Pharm 2012; 436:194-200.
24. Geetha V. S, Malarkodi Velraj. Formulation, Optimization And In Vitro Evaluation Of 5-Fluorouracil Loaded Liquorice Crude Protein Nanoparticles For Sustained Drug Delivery Using Boxbehnken Design. Int J App Pharm. 2021;13:216-26.
25. Dash S, Murthy PN, Nath L, Chowdhury P. Kinetic modeling on drug release from controlled drug delivery systems. Acta Pol Pharm 2010; 67:217-23.
This work is licensed under a Creative Commons Attribution 4.0 International License.