INNOVATIVE LIPIDIC NANOCARRIERS OF FLUTAMIDE ENHANCING ITS IN VITRO CYTOTOXICITY AND IN VIVO ORAL BIOAVAILABILITY: DESIGN, OPTIMIZATION, CHARACTERIZATION, AND PHARMACOKINETIC ASPECTS

MOHAMED A. ALI; MAGDY I. MOHAMED; KHALID M. EL-SAY; MOHAMED A. MEGAHED

doi:10.22159/ijap.2024v16i4.51037

Authors

MOHAMED A. ALI Department of Pharmaceutics and Industrial Pharmacy, Faculty of Pharmacy, Cairo University, Cairo, Egypt
MAGDY I. MOHAMED Department of Pharmaceutics and Industrial Pharmacy, Faculty of Pharmacy, Cairo University, Cairo, Egypt https://orcid.org/0000-0002-0480-7720
KHALID M. EL-SAY Department of Pharmaceutics, Faculty of Pharmacy, King Abdulaziz University, Jeddah-21589, Saudi Arabia https://orcid.org/0000-0002-5539-3193
MOHAMED A. MEGAHED Department of Pharmaceutics and Pharmaceutical Technology, Egyptian Russian University, Cairo, Egypt https://orcid.org/0000-0001-5887-4235

DOI:

https://doi.org/10.22159/ijap.2024v16i4.51037

Keywords:

Bioavailability, Box-behnken design, Flutamide, PC-3 cell line, MTT assay, Optimization, Pharmacokinetic

Abstract

Objective: the reduced oral bioavailability of Flutamide has hindered its effectiveness as a chemotherapeutic agent for prostate cancer treatment. Our study aimed to enhance FLUTAMIDE in vitro cytotoxicity and oral bioavailability via its incorporation into lipid nanocarriers that contained solid lipid (Precirol^®) alone or in combination with anti-androgenic oils such as Saw Palmetto Oil (SPO) and Pumpkin Seed Oil (PSO).

Methods: we employed the Box Behnken Design (BBD) to optimize Flutamide-loaded nanocarriers, focusing on mean vesicular size, zeta potential, and entrapment efficiency.

Results: the optimized nanovesicles exhibited dimensions of 330.2 nm, a zeta potential of -43.1 mV, and an entrapment efficiency of 66.1%. Morphological analysis using Transition Electron Microscope (TEM) and Scanning Electron Microscope (SEM) confirmed the spherical shape of the nanovesicles. Differntial Scanning Calorimetry (DSC) thermograms and X-ray diffractograms indicated decreased crystallinity of encapsulated Flutamide compared to free Flutamide. In vitro cytotoxicity studies demonstrated enhanced effects against prostate cancer cells (PC-3) for optimized Flutamide-loaded nanocarriers containing the 2 anti-androgenic oils over both nanocarriers containing no oils and free Flutamide suspension. In vivo pharmacokinetic analysis in male rats showed increased oral bioavailability for flutamide-loaded nanocarriers with C_max values of 559.35±41.79 ng/ml and 670.9±24.61 ng/ml for different formulations compared to the free flutamide suspension with a C_max value of 281.4±94.33 ng/ml.

Conclusion: These findings support FLUTAMIDE oral bioavailability improvement through nanocarriers encapsulation, advocating its utilization in prostate cancer therapy and approving the additive anti-androgenic effect after its combination with SPO and PSO.

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References

Blair HA. Hexanic extract of serenoa repens (Permixon®): a review in symptomatic benign prostatic hyperplasia. Drugs Aging. 2022;39(3):235-43. doi: 10.1007/s40266-022-00924-3, PMID 35237936.

leisegang K, Jimenez M, Durairajanayagam D, Finelli R, Majzoub A, Henkel R. Phytomedicine plus a systematic review of herbal medicine in the clinical treatment of benign prostatic hyperplasia. Phytomedicine Plus. 2022;2(1):100-53. doi: 10.1016/j.phyplu.2021.100153.

Ali MA, Mohamed MI, Megahed MA, Abdelghany TM, El-Say KM. Cholesterol-based nanovesicles enhance the in vitro cytotoxicity, ex vivo intestinal absorption, and in vivo bioavailability of flutamide. Pharmaceutics. 2021;13(11):1-22. doi: 10.3390/pharmaceutics13111741, PMID 34834155.

Giorgetti R, di Muzio M, Giorgetti A, Girolami D, Borgia L, Tagliabracci A. Flutamide-induced hepatotoxicity: ethical and scientific issues. Eur Rev Med Pharmacol Sci. 2017;21(1)Suppl:69-77. PMID 28379593.

Khan N, Abdelhamid HN, Yan JY, Chung FT, Wu HF. Detection of flutamide in pharmaceutical dosage using higher electrospray ionization mass spectrometry (ESI-MS) tandem mass coupled with Soxhlet apparatus. Anal Chem Res. 2015;3(Feb):89-97. doi: 10.1016/j.ancr.2015.01.001.

Rojas PA, Iglesias TG, Barrera F, Mendez GP, Torres J, San Francisco IF. Acute liver failure and liver transplantation secondary to flutamide treatment in a prostate cancer patient. Urol Case Rep. 2020;33:101370. doi: 10.1016/j.eucr.2020.101370, PMID 33102069.

Jeevana JB, Sreelakshmi K. Design and evaluation of self-nanoemulsifying drug delivery system of flutamide. J Young Pharm. 2011;3(1):4-8. doi: 10.4103/0975-1483.76413, PMID 21607048.

Elgindy N, Elkhodairy K, Molokhia A, Elzoghby A. Lyophilization monophase solution technique for improvement of the physicochemical properties of an anticancer drug, flutamide. Eur J Pharm Biopharm. 2010;74(2):397-405. doi: 10.1016/j.ejpb.2009.11.011, PMID 19944160.

Youssef SF, Elnaggar YS, Abdallah OY. Elaboration of polymersomes versus conventional liposomes for improving oral bioavailability of the anticancer flutamide. Nanomedicine (Lond). 2018;13(23):3025-36. doi: 10.2217/nnm-2018-0238, PMID 30507344.

Bakhaidar RB, Hosny KM, Mahier IM, Rizq WY, Safhi AY, Bukhary DM. Development and optimization of a tamsulosin nanostructured lipid carrier loaded with saw palmetto oil and pumpkin seed oil for treatment of benign prostatic hyperplasia. Drug Deliv. 2022;29(1):2579-91. doi: 10.1080/10717544.2022.2105448, PMID 35915055.

Andjelkovic M, Van Camp J, Trawka A, Verhe R. Phenolic compounds and some quality parameters of pumpkin seed oil. Euro J Lipid Sci & Tech. 2010;112(2):208-17. doi: 10.1002/ejlt.200900021.

Alhakamy NA, Fahmy UA, Ahmed OA. Attenuation of benign prostatic hyperplasia by optimized tadalafil loaded pumpkin seed oil-based self nanoemulsion: in vitro and in vivo evaluation. Pharmaceutics. 2019;11(12):1-13. doi: 10.3390/pharmaceutics11120640, PMID 31805693.

Csikos E, Horvath A, Acs K, Papp N, Balazs VL, Dolenc MS. Treatment of benign prostatic hyperplasia by natural drugs. Molecules. 2021;26(23):1-32. doi: 10.3390/molecules26237141, PMID 34885733.

Anastassakis K. Androgenetic alopecia from A to Z. 1st ed. Vol. 2, Androgenetic Alopecia From A to Z. Springer Cham; 2022. p. 193-7.

Kalwat JI. The use of serenoa repens (saw palmetto) in hair care products. BJSTR. 2019;13(1):9725-8. doi: 10.26717/BJSTR.2019.13.002348.

Zhang Y, Zhang P, Zhu T. Ovarian carcinoma biological nanotherapy: comparison of the advantages and drawbacks of lipid, polymeric, and hybrid nanoparticles for cisplatin delivery. Biomed Pharmacother. 2019;109(1):475-83. doi: 10.1016/j.biopha.2018.10.158, PMID 30399584.

Davis ME, Chen ZG, Shin DM. Nanoparticle therapeutics: an emerging treatment modality for cancer. Nat Rev Drug Discov. 2008;7(9):771-82. doi: 10.1038/nrd2614, PMID 18758474.

Xin Y, Huang Q, Tang JQ, Hou XY, Zhang P, Zhang LZ. Nanoscale drug delivery for targeted chemotherapy. Cancer Lett. 2016;379(1):24-31. doi: 10.1016/j.canlet.2016.05.023, PMID 27235607.

Nguyen CN, Nguyen TT, Nguyen HT, Tran TH. Nanostructured lipid carriers to enhance transdermal delivery and efficacy of diclofenac. Drug Deliv Transl Res. 2017;7(5):664-73. doi: 10.1007/s13346-017-0415-2, PMID 28776220.

Carbone C, Campisi A, Musumeci T, Raciti G, Bonfanti R, Puglisi G. FA-loaded lipid drug delivery systems: preparation, characterization and biological studies. Eur J Pharm Sci. 2014;52(1):12-20. doi: 10.1016/j.ejps.2013.10.003, PMID 24514450.

Vidlarova L, Hanus J, Vesely M, Ulbrich P, Stepanek F, Zbytovska J. Effect of lipid nanoparticle formulations on skin delivery of a lipophilic substance. Eur J Pharm Biopharm. 2016;108:289-96. doi: 10.1016/j.ejpb.2016.07.016, PMID 27449632.

Muller RH, Radtke M, Wissing SA. Nanostructured lipid matrices for improved microencapsulation of drugs. Int J Pharm. 2002;242(1-2):121-8. doi: 10.1016/s0378-5173(02)00180-1, PMID 12176234.

Üner M. Preparation, characterization and physico-chemical properties of solid lipid nanoparticles (SLN) and nanostructured lipid carriers (NLC): their benefits as colloidal drug carrier systems. Pharmazie. 2006;61(5):375-86. PMID 16724531.

Aslam R, Tiwari V, Upadhyay P, Tiwari A. Original article revolutionizing therapeutic delivery: diosgenin-loaded solid lipid nanoparticles unleash advanced carriers. International Journal of Applied Pharmaceutics. 2024;16(1):124–33. doi: 10.22159/ijap.2024v16i1.49306.

lópez KL, Ravasio A, Gonzalez Aramundiz JV, Zacconi FC. Solid lipid nanoparticles (SLN) and nanostructured lipid carriers (NLC) prepared by microwave and ultrasound-assisted synthesis: promising green strategies for the nanoworld. Pharmaceutics. 2023;15(5):1-26. doi: 10.3390/pharmaceutics15051333, PMID 37242575.

El-Say KM, Hosny KM. Optimization of carvedilol solid lipid nanoparticles: an approach to control the release and enhance the oral bioavailability on rabbits. PLOS ONE. 2018;13(8):e0203405. doi: 10.1371/journal.pone.0203405, PMID 30161251.

El-Say KM, El-Helw AR, Ahmed OA, Hosny KM, Ahmed TA, Kharshoum RM. Statistical optimization of controlled release microspheres containing cetirizine hydrochloride as a model for water-soluble drugs. Pharm Dev Technol. 2015;20(6):738-46. doi: 10.3109/10837450.2014.920353, PMID 24856961.

Kudarha R, Dhas NL, Pandey A, Belgamwar VS, Ige PP. Box-behnken study design for optimization of bicalutamide-loaded nanostructured lipid carrier: stability assessment. Pharm Dev Technol. 2015;20(5):608-18. doi: 10.3109/10837450.2014.908305, PMID 24785784.

Ferreira N, Viana T, Henriques B, Tavares DS, Jacinto J, Colonia J. Application of response surface methodology and box-behnken design for the optimization of mercury removal by Ulva sp J Hazard Mater. 2023;445:130405. doi: 10.1016/j.jhazmat.2022.130405, PMID 36437192.

Aodah AH, Hashmi S, Akhtar N, Ullah Z, Zafar A, Zaki RM. Formulation development, optimization by box-behnken design, and in vitro and ex vivo characterization of hexatriacontane-loaded transethosomal gel for antimicrobial treatment for skin infections. Gels. 2023;9(4):1-22. doi: 10.3390/gels9040322, PMID 37102934.

Patra S. Development, characterization and pharmacokinetic evaluation of optimized vildagliptin sustained release matrix tablet using box-behnken design. International Journal of Applied Pharmaceutics. 2024;16(1):214-23. doi: 10.22159/ijap.2024v16i1.48052.

Hosny KM, Aljaeid BM. Sildenafil citrate as oral solid lipid nanoparticles: a novel formula with higher bioavailability and sustained action for treatment of erectile dysfunction. Expert Opin Drug Deliv. 2014;11(7):1015-22. doi: 10.1517/17425247.2014.912212, PMID 24746063.

Li Y, Zhao X, Zu Y, Zhang Y. Preparation and characterization of paclitaxel nanosuspension using novel emulsification method by combining high speed homogenizer and high pressure homogenization. Int J Pharm. 2015;490(1-2):324-33. doi: 10.1016/j.ijpharm.2015.05.070, PMID 26027492.

Ahmed TA, El-Say KM, Aljaeid BM, Fahmy UA, Abd-Allah FI. Transdermal glimepiride delivery system based on optimized ethosomal nano-vesicles: Preparation, characterization, in vitro, ex vivo and clinical evaluation. Int J Pharm. 2016;500(1-2):245-54. doi: 10.1016/j.ijpharm.2016.01.017, PMID 26775063.

Manconi M, Sinico C, Valenti D, lai F, Fadda AM. Niosomes as carriers for tretinoin. III. A study into the in vitro cutaneous delivery of vesicle-incorporated tretinoin. Int J Pharm. 2006;311(1-2):11-9. doi: 10.1016/j.ijpharm.2005.11.045, PMID 16439071.

El-Ridy MS, Abd ElRahman AA, Awad GM, Khalil RM, Younis MM. In vitro and in vivo evaluation of niosomes containing celecoxib. IJPSR. 2014;5(11):4677-88. doi: 10.13040/IJPSR.0975-8232.5(11).4677-88.

Elzoghby AO, Helmy MW, Samy WM, Elgindy NA. Novel ionically crosslinked casein nanoparticles for flutamide delivery: formulation, characterization, and in vivo pharmacokinetics. Int J Nanomedicine. 2013;8:1721-32. doi: 10.2147/IJN.S40674, PMID 23658490.

Mosmann T. Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. J Immunol Methods. 1983;65(1-2):55-63. doi: 10.1016/0022-1759(83)90303-4, PMID 6606682.

Scudiero DA, Shoemaker RH, Paull KD, Monks A, Tierney S, Nofziger TH. Evaluation of a soluble tetrazolium/formazan assay for cell growth and drug sensitivity in culture using human and other tumor cell lines. Cancer Res. 1988;48(17):4827-33. PMID 3409223.

Elzoghby AO, Helmy MW, Samy WM, Elgindy NA. Spray-dried casein-based micelles as a vehicle for solubilization and controlled delivery of flutamide: formulation, characterization, and in vivo pharmacokinetics. Eur J Pharm Biopharm. 2013;84(3):487-96. doi: 10.1016/j.ejpb.2013.01.005, PMID 23403015.

Khames A, Khaleel MA, El-Badawy MF, El-Nezhawy AO. Natamycin solid lipid nanoparticles-sustained ocular delivery system of higher corneal penetration against deep fungal keratitis: preparation and optimization. Int J Nanomedicine. 2019;14:2515-31. doi: 10.2147/IJN.S190502, PMID 31040672.

Madhusudhan B, Rambhau D, Apte SS, Gopinath D. Oral bioavailability of flutamide from 1-o-alkylglycerol stabilized o/w nanoemulsions. J Dispers Sci Technol. 2007;28(8):1254-61. doi: 10.1080/01932690701528241.

Chalikwar SS, Belgamwar VS, Talele VR, Surana SJ, Patil MU. Formulation and evaluation of nimodipine-loaded solid lipid nanoparticles delivered via lymphatic transport system. Colloids Surf B Biointerfaces. 2012;97:109-16. doi: 10.1016/j.colsurfb.2012.04.027, PMID 22609590.

Araujo J, Gonzalez Mira E, Egea MA, Garcia ML, Souto EB. Optimization and physicochemical characterization of a triamcinolone acetonide-loaded NLC for ocular antiangiogenic applications. Int J Pharm. 2010;393(1-2):167-75. doi: 10.1016/j.ijpharm.2010.03.034, PMID 20362042.

Kumbhar DD, Pokharkar VB. Engineering of a nanostructured lipid carrier for the poorly water-soluble drug, bicalutamide: physicochemical investigations. Colloids and Surfaces A: Physicochemical and Engineering Aspects. 2013;416:32-42. doi: 10.1016/j.colsurfa.2012.10.031.

Walbi IA, Ahmad MZ, Ahmad J, Algahtani MS, Alali AS, Alsudir SA. Development of a curcumin-loaded lecithin/chitosan nanoparticle utilizing a box-behnken design of experiment: formulation design and influence of process parameters. Polymers (Basel). 2022;14(18):1-17. doi: 10.3390/polym14183758, PMID 36145903.

Agrawal Y, Petkar KC, Sawant KK. Development, evaluation and clinical studies of acitretin-loaded nanostructured lipid carriers for topical treatment of psoriasis. Int J Pharm. 2010;401(1-2):93-102. doi: 10.1016/j.ijpharm.2010.09.007, PMID 20858539.

WC. Ageing effects in parenteral fat emulsions: the role of fatty acids. International Journal of Pharmaceutics. 1987;39:33-7.

Chinsriwongkul A, Chareanputtakhun P, Ngawhirunpat T, Rojanarata T, Sila-on W, Ruktanonchai U. Nanostructured lipid carriers (NLC) for parenteral delivery of an anticancer drug. AAPS PharmSciTech. 2012;13(1):150-8. doi: 10.1208/s12249-011-9733-8, PMID 22167418.

Guada M, Sebastian V, Irusta S, Feijoo E, Dios Vieitez Mdel C, Blanco Prieto MJ. Lipid nanoparticles for cyclosporine a administration: development, characterization, and in vitro evaluation of their immunosuppression activity. Int J Nanomedicine. 2015;10:6541-53. doi: 10.2147/IJN.S90849, PMID 26527872.

Aboud HM, El Komy MH, Ali AA, El Menshawe SF, Abd Elbary A. Development, optimization, and evaluation of carvedilol-loaded solid lipid nanoparticles for intranasal drug delivery. AAPS PharmSciTech. 2016;17(6):1353-65. doi: 10.1208/s12249-015-0440-8, PMID 26743643.

Bondì ML, Craparo EF, Giammona G, Cervello M, Azzolina A, Diana P. Nanostructured lipid carriers-containing anticancer compounds: preparation, characterization, and cytotoxicity studies. Drug Deliv. 2007;14(2):61-7. doi: 10.1080/10717540600739914, PMID 17364869.

Chandratre SS, Dash AK. Multifunctional nanoparticles for prostate cancer therapy. AAPS PharmSciTech. 2015;16(1):98-107. doi: 10.1208/s12249-014-0202-z, PMID 25190362.

Mortazavi SM, Mohammadabadi MR, Khosravi Darani K, Mozafari MR. Preparation of liposomal gene therapy vectors by a scalable method without using volatile solvents or detergents. J Biotechnol. 2007;129(4):604-13. doi: 10.1016/j.jbiotec.2007.02.005, PMID 17353061.

Shenoy VS, Gude RP, Murthy RS. In vitro anticancer evaluation of 5-fluorouracil lipid nanoparticles using B16F10 melanoma cell lines. Int Nano Lett. 2013;3(1):36. doi: 10.1186/2228-5326-3-36.

Tavano L, Aiello R, Ioele G, Picci N, Muzzalupo R. Niosomes from glucuronic acid-based surfactant as new carriers for cancer therapy: preparation, characterization and biological properties. Colloids Surf B Biointerfaces. 2014;118:7-13. doi: 10.1016/j.colsurfb.2014.03.016, PMID 24709252.

Danhier F, lecouturier N, Vroman B, Jerome C, Marchand Brynaert J, Feron O. Paclitaxel-loaded pegylated PLGA-based nanoparticles: in vitro and in vivo evaluation. J Control Release. 2009;133(1):11-7. doi: 10.1016/j.jconrel.2008.09.086, PMID 18950666.

Sharma V, Anandhakumar S, Sasidharan M. Self-degrading niosomes for encapsulation of hydrophilic and hydrophobic drugs: an efficient carrier for cancer multi-drug delivery. Mater Sci Eng C Mater Biol Appl. 2015 Nov;56:393-400. doi: 10.1016/j.msec.2015.06.049, PMID 26249606.

Dolatabadi S, Karimi M, Nasirizadeh S, Hatamipour M, Golmohammadzadeh S, Jaafari MR. Preparation, characterization and in vivo pharmacokinetic evaluation of curcuminoids-loaded solid lipid nanoparticles (SLNs) and nanostructured lipid carriers (NLCs). J Drug Deliv Sci Technol. 2021;62:102352. doi: 10.1016/j.jddst.2021.102352.

Veerareddy PR, Bobbala SK. Enhanced oral bioavailability of isradipine via proniosomal systems. Drug Dev Ind Pharm. 2013;39(6):909-17. doi: 10.3109/03639045.2012.717945, PMID 22998221.