Screening of the most efficient variables for the development of protein nanoparticles for anticancer drug by the design of the experiment

Mustafa Egla; Nawal Rajab

Screening of the most efficient variables for the development of protein nanoparticles for anticancer drug by the design of the experiment

Abstract

The goal of the current research was to apply the design of the experiment (DoE) method "Taguchi orthogonal array (OA) model" to fabricate sericin-based nanoparticles (NPs) of paclitaxel (PTX). The DoE aimed to examine the influence of seven factors on responses, including particle size, polydispersity index (PDI), and percent of entrapment efficiency (EE%). The factors include the concentrations of (protein, poloxamer 407, tween 80), stirring (speed and time), the rate of addition of organic solvent, and finally, the ratio of organic to aqueous solvents. NPs were made using the desolvation method. The NPs were investigated for particle size, PDI, and EE%, then optimized utilizing the desirability function. The optimized formulation with higher desirability was validated by comparing the predicted properties with the experimental formulation. After that, it was tested for in vitro release study, morphological study by field emission scanning electron microscope (FESEM), and zeta potential. The results showed poloxamer 407 and sericin concentrations constituted this study's two most influential variables on particle size. For PDI, the most influential parameter was sericin concentration. Stirring speed was the most influential parameter on EE%. The formulation 6 produced a high desirability function (0.83). The validation procedure indicated a minor variation between actual and predicted data, indicating a high connection between experimental and predicted data. These NPs release PTX slowly. It possessed enough surface charge to be stable. In conclusion, our innovative systematic method optimizes and evaluates nanocarriers with a few clever trials.

Keywords

References

[1] Wang F, Porter M, Konstantopoulos A, Zhang P, Cui H. Preclinical development of drug delivery systems for paclitaxel-based cancer chemotherapy. J Control Release. 2017;267:100-118. https://doi.org/10.1016/j.jconrel.2017.09.026
[2] Liu Y, Zhang Y, Yan Q, Zhong X, Hu C. Evaluation of microstructure, dissolution rate, and oral bioavailability of paclitaxel poloxamer 188 solid dispersion. Drug Deliv Transl Res. 2024;14(2):329-341. https://doi.org/10.1007/s13346-023-01400-0
[3] Hussien RM, Ghareeb MM. Formulation and characterization of isradipine nanoparticle for dissolution enhancement. Iraqi J Pharm Sci. 2021;30(1):218-225. https://doi.org/10.31351/vol30iss1pp218-225
[4] Nugroho AK, Kusumorini N, Pramono S, Martien R. An update on nanoparticle formulation design of piperine to improve its oral bioavailability: A Review. Iraqi J Pharm Sci. 2023;32(1):14-30. https://doi.org/10.31351/vol32iss1pp14-30
[5] Elzoghby AO, Samy WM, Elgindy NA. Protein-based nanocarriers as promising drug and gene delivery systems. J Control Release. 2012;161(1):38-49. https://doi.org/10.1016/j.jconrel.2012.04.036
[6] Muhesen RA, Rajab NA. Formulation and characterization of olmesartan medoxomil as a nanoparticle. Res J Pharm Technol . 2023;16(7):3314-3320. http://dx.doi.org/10.52711/0974-360X.2023.00547
[7] Alkawak RSY, Rajab NA. Lornoxicam-loaded cubosomes:-Preparation and in vitro characterization. Iraqi J Pharm Sci. 2022;31(1):144-153. https://doi.org/10.31351/vol31iss1pp144-153
[8] Khosropanah MH, Vaghasloo MA, Shakibaei M, Mueller AL, Kajbafzadeh AM, Amani L, Haririan I, Azimzadeh A, Hassannejad Z, Zolbin MM. Biomedical applications of silkworm (Bombyx Mori) proteins in regenerative medicine (a narrative review). J Tissue Eng Regen Med. 2022;16(2):91-109. https://doi.org/10.1002/term.3267

[9] Altememy D, Khoobi M, Javar HA, Alsamarrai S, Khosravian P. Synthesis and characterization of silk fibroin-coated mesoporous silica nanoparticles for tioguanine targeting to leukemia. Int J Pharm Res. 2020;12(supp2):1181-1192. http://dx.doi.org/10.31838/ijpr/2020.SP2.145
[10] Mahalingam M, Krishnamoorthy K. Fabrication and optimization of camptothecin loaded Eudragit S 100 nanoparticles by Taguchi L4 orthogonal array design. Int J Pharm Investig. 2015 5(3):147-154. https://doi.org/10.4103%2F2230-973X.160852
[11] Do Kim K, Choi DW, Choa Y-H, Kim HT. Optimization of parameters for the synthesis of zinc oxide nanoparticles by Taguchi robust design method. Colloids Surf A: Physicochem Eng Asp. 2007;311(1-3):170-173. http://dx.doi.org/10.1016/j.colsurfa.2007.06.017
[12] Torkaman R, Soltanieh M, Kazemian H. Optimization of parameters for synthesis of MFI nanoparticles by Taguchi robust design. Chem Eng Technol. 2010;33(6):902-910. http://dx.doi.org/10.1002/ceat.200900367
[13] Rampado R, Peer D. Design of experiments in the optimization of nanoparticle-based drug delivery systems. J Control Release. 2023;358:398-419. https://doi.org/10.1016/j.jconrel.2023.05.001
[14] Kanoujia J, Faizan M, Parashar P, Singh N, Saraf SA. Curcumin loaded sericin nanoparticles: Assessment for biomedical application. Food Hydrocoll Health. 2021;1:100029. https://doi.org/10.1016/j.fhfh.2021.100029
[15] Syed A, Devi VK, Karwa P. Applying Taguchi design for the synthesis of albumin nanoparticles of methotrexate by desolvation technique. RGUHS J Pharm Sci. 2020;10(4):39-44.
[16] Kharia AA, Singhai AK. Effective parameters for formulation of gastro adhesive nanoparticles: Screening by design-of-experiments approach. J Microencapsul. 2014;31(4):399-405. https://doi.org/10.3109/02652048.2013.863398.
[17] Pandey AP, More MP, Karande KP, Chitalkar RV, Patil PO, Deshmukh PK. Optimization of desolvation process for fabrication of lactoferrin nanoparticles using quality by design approach. Artif Cells Nanomed Biotechnol. 2017;45(6):1-14. https://doi.org/10.1080/21691401.2016.1202259
[18] Lee SH, Heng D, Ng WK, Chan HK, Tan RB. Nano spray drying: a novel method for preparing protein nanoparticles for protein therapy. Int J Pharm. 2011;403(1-2):192-200. https://doi.org/10.1016/j.ijpharm.2010.10.012
[19] Pham D-D, Fattal E, Tsapis N. Pyrazinamide-loaded poly (lactide-co-glycolide) nanoparticles: Optimization by experimental design. J Drug Deliv Sci Technol. 2015;30:384-390. https://doi.org/10.1016/j.jddst.2015.07.006
[20] Esim O, Gedik ME, Dogan AL, Gunaydin G, Hascicek C. Development of carboplatin loaded bovine serum albumin nanoparticles and evaluation of its effect on an ovarian cancer cell line. J Drug Deliv Sci Technol. 2021;64:102655. https://doi.org/10.1016/j.jddst.2021.102655
[21] Hassanzadeh S, Nematollahzadeh A, Mirzayi B, Kaboli SF. Protein-based nanoparticles synthesized at a high shear rate and optimized for drug delivery applications. J Mol Liq. 2021;335:116133. http://dx.doi.org/10.1016/j.molliq.2021.116133
[22] Hashim AA-J, Rajab NA. Anastrozole loaded nanostructured lipid carriers: Preparation and evaluation. Iraqi J Pharm Sci. 2021;30(2):185-195. https://doi.org/10.31351/vol30iss2pp185-195
[23] Suktham K, Koobkokkruad T, Wutikhun T, Surassmo S. Efficiency of resveratrol-loaded sericin nanoparticles: Promising bionanocarriers for drug delivery. Int J Pharm. 2018;537(1-2):48-56. https://doi.org/10.1016/j.ijpharm.2017.12.015
[24] Jassem NA, Rajab NA. Formulation and in vitro evaluation of azilsartan medoxomil nanosuspension. Int J Pharm Pharm Sci. 2017;9(7):110-119. http://dx.doi.org/10.22159/ijpps.2017v9i7.18917
[25] Al-Sawahli MM, Zarka MA, Ghareeb MM, Kutbi HI, Kammoun AK, Noreddin AM, El-Telbany DFA. Zein nanoplatforms for functional promotion of pro-apoptosis and antiproliferation induced in human breast carcinoma MCF7 cells by pterostilbene. Perspect Recent Adv Med Res. 2023; 12: 93-114. https://doi.org/10.9734/bpi/pramr/v12/9322F
[26] Kouchakinejad R, Shariati S, Abolhasani J, Kalhor EG, Vardini MT. Core-shells of magnetite nanoparticles decorated by SBA-3-SO3H mesoporous silica for magnetic solid phase adsorption of paraquat herbicide from aqueous solutions. Colloids Surf A Physicochem Eng Asp. 2022;643:128709. http://dx.doi.org/10.1016/j.colsurfa.2022.128709
[27] Schubert MA, Müller-Goymann CC. Solvent injection as a new approach for manufacturing lipid nanoparticles--evaluation of the method and process parameters. Eur J Pharm Biopharm. 2003;55(1):125-131. https://doi.org/10.1016/s0939-6411(02)00130-3
[28] Asasutjarit R, Sorrachaitawatwong C, Tipchuwong N, Pouthai S. Effect of formulation compositions on particle size and zeta potential of diclofenac sodium-loaded chitosan nanoparticles. Int J Pharm Pharm Sci. 2013;7(9):568-570. doi.org/10.5281/zenodo.1087714
[29] Saberi AH, Fang Y, McClements DJ. Fabrication of vitamin E-enriched nanoemulsions: factors affecting particle size using spontaneous emulsification. J Colloid Interface Sci. 2013;391:95-102. https://doi.org/10.1016/j.jcis.2012.08.069
[30] Xiao F, Hu X, Chen Y, Zhang Y. Porous Zr-based metal-organic frameworks (Zr-MOFs)-ıncorporated thin-film nanocomposite membrane toward enhanced desalination performance. ACS Appl Mater Interfaces. 2019;11(50):47390-47403. https://doi.org/10.1021/acsami.9b17212.
[31] Felton LA. Mechanisms of polymeric film formation. Int J Pharm. 2013;457(2):423-427. https://doi.org/10.1016/j.ijpharm.2012.12.027
[32] Abedinpour N, Ghanbariasad A, Taghinezhad A, Osanloo M. Preparation of nanoemulsions of mentha piperita essential oil and investigation of their cytotoxic effect on human breast cancer lines. BioNanoScience. 2021;11:428-436. https://doi.org/10.1007/s12668-021-00827-4
[33] Li X, Li Y, Hua Y, Qiu A, Yang C, Cui S. Effect of concentration, ionic strength and freeze-drying on the heat-induced aggregation of soy proteins. Food Chem. 2007;104(4):1410-1417. http://dx.doi.org/10.1016/j.foodchem.2007.02.003
[34] Galisteo-Gonzalez F, Molina-Bolivar J. Systematic study on the preparation of BSA nanoparticles. Colloids Surf B Biointerfaces. 2014;123:286-292. https://doi.org/10.1016/j.colsurfb.2014.09.028
[35] Usama A, Fetih G, El-Faham T. Performance of meloxicam niosomal gel formulations for transdermal drug delivery. J Pharm Res Int. 2016;12(2):1-14. https://doi.org/10.9734/BJPR/2016/26985
[36] Li L, Kim JK, Huh KM, Lee Y-k, Kim SY. Targeted delivery of paclitaxel using folate-conjugated heparin-poly (ƒÀ-benzyl-l-aspartate) self-assembled nanoparticles. Carbohydr Polym. 2012;87(3):2120-2108. http://dx.doi.org/10.1016/j.carbpol.2011.10.030
[37] Sallam NM, Sanad RAB, Ahmed MM, Khafagy ES, Ghorab M, Gad S. Impact of the mucoadhesive lyophilized wafer loaded with novel carvedilol nano-spanlastics on biochemical markers in the heart of spontaneously hypertensive rat models. Drug Deliv Transl Res. 2021;11:1009-1036. https://doi.org/10.1007/s13346-020-00814-4
[38] Paranthaman S, Uthaiah CA, Osmani RAM, Hani U, Ghazwani M, Alamri AH, Fatease AA, Madhunapantula SV, Gowda DV. Anti-proliferative potential of quercetin loaded polymeric mixed micelles on rat C6 and human U87MG glioma cells. Pharmaceutics. 2022;14(8):1643. https://doi.org/10.3390/pharmaceutics14081643
[39] Dinh L, Hong J, Kim DM, Lee G, Park EJ, Baik SH, Hwang S-J. A novel thermosensitive poloxamer-hyaluronic acid-kappa-carrageenan-based hydrogel anti-adhesive agent loaded with 5-fluorouracil: A preclinical study in Sprague-Dawley rats. Int J Pharm. 2022;621:121771. https://doi.org/10.1016/j.ijpharm.2022.121771
[40] Sanjula B, Shah FM, Javed A, Alka A. Effect of poloxamer 188 on lymphatic uptake of carvedilol-loaded solid lipid nanoparticles for bioavailability enhancement. J Drug Target. 2009;17(3):249-256. https://doi.org/10.1080/10611860902718672
[41] Cai H, Xiang Y, Zeng Y, Li Z, Zheng X, Luo Q, Zhu H, Gong Q, Gu Z, Liu Y. Cathepsin B-responsive and gadolinium-labeled branched glycopolymer-PTX conjugate-derived nanotheranostics for cancer treatment. Acta Pharm Sin B. 2021;11(2):544-559. https://doi.org/10.1016/j.apsb.2020.07.023
[42] Qu G, Hou S, Qu D, Tian C, Zhu J, Xue L, Ju C, Zhang C. Self-assembled micelles based on N-octyl-Nf-phthalyl-O-phosphoryl chitosan derivative as an effective oral carrier of paclitaxel. Carbohydr Polym. 2019;207:428-439. https://doi.org/10.1016/j.carbpol.2018.11.099
[43] Carissimi G, Lozano-Perez AA, Montalban MG, Aznar-Cervantes SD, Cenis JL, Villora G. Revealing the influence of the degumming process in the properties of silk fibroin nanoparticles. Polymers. 2019;11(12):2045. https://doi.org/10.3390%2Fpolym11122045
[44] Pedro IDR, Almeida OP, Martins HR, de Alcantara Lemos J, de Barros ALB, Leite EA, Carneiro G. Optimization and in vitro/in vivo performance of paclitaxel-loaded nanostructured lipid carriers for breast cancer treatment. J Drug Deliv Sci Technol . 2019;54:101370. https://doi.org/10.1016/j.jddst.2019.101370
[45] Alkufi HK, Kassab HJ. Formulation and evaluation of sustained release sumatriptan mucoadhesive intranasal in-situ gel. Iraqi J Pharm Sci. 2019;28(2):95-104. https://doi.org/10.31351/vol28iss2pp95-104
[46] Saha J, Mondal M, Sheikh MK, Habib M. Extraction, structural and functional properties of silk sericin biopolymer from Bombyx mori silk cocoon waste. J Text Sci Eng. 2019;9(1): 1000390. http://dx.doi.org/10.4172/2165-8064.1000390
[47] Jangid AK, Pooja D, Jain P, Gupta N, Ramesan S, Kulhari H. Self-assembled and pH-responsive polymeric nanomicelles impart effective delivery of paclitaxel to cancer cells. RSC Adv. 2021;11(23):13928-13939. https://doi.org/10.1039%2Fd1ra01574e
[48] AlKhalidi MM, Jawad FJ. Enhancement of aqueous solubility and dissolution rate of etoricoxib by solid dispersion technique. Iraqi J Pharm Sci. 2020;29(1):76-87. https://doi.org/10.31351/vol29iss1pp76-87
[49] Balcao VM, Harada LK, Jorge LR, Oliveira Jr JM, Tubino M, Vila MM. Structural and functional stabilization of sericin from Bombyx mori cocoons in a biopolysaccharide film: Bioorigami for skin regeneration. J Braz.l Chem Soc. 2020;31:833-848. https://doi.org/10.21577/0103-5053.20190247
[50] Egla M, Hammid S. Design zolmitriptan liquisolid orodispersible tablets and their in vitro evaluation. J Chem Pharm Res. 2016;8(11):232-242. http://dx.doi.org/10.22159/ijpps.2017v9i1.15656
[51] Al-Sawaf OF, Jalal F. Novel probe sonication method for the preparation of meloxicam bilosomes for transdermal delivery: Part One. J Res Med Dent Sci. 2023;11(6):5-12.
[52] Khattak MA, Iqbal Z, Fazli N, Khan FU, Khan S, Gohar S, Sakhi M, Iqbal M, Pervez S. Using the design of experiments for development, validation, and optimization of HPLC-UV assay of tamoxifen citrate in polymeric nanoparticles. Pharm Chem J. 2022;56(9):1297-1306. http://dx.doi.org/10.1007/s11094-022-02789-7
[53] Taheri A, Razavi SM. Fabrication of cress seed gum nanoparticles, an anionic polysaccharide, using desolvation technique: An optimization study. BioNanoScience. 2015;5:104-116. http://dx.doi.org/10.1007/s12668-015-0169-6
[54] Orlandi G, Bari E, Catenacci L, Sorrenti M, Segale L, Farago S, Sorlini M, Arciola CR, Torre ML, Perteghella S. Polyphenols-loaded sericin self-assembling nanoparticles: A slow-release for regeneration by tissue-resident mesenchymal stem/stromal cells. Pharmaceutics. 2020;12(4):381. https://doi.org/10.3390/pharmaceutics12040381
[55] Mandal BB, Kundu S. Self-assembled silk sericin/poloxamer nanoparticles as nanocarriers of hydrophobic and hydrophilic drugs for targeted delivery. Nanotechnology. 2009;20(35):355101. http://dx.doi.org/10.1088/0957-4484/20/35/355101
[56] Ghazy E, Abdulrasool AA, Al-Tamimi JJ, Ayash N. Nebivolol hydrochloride loaded nanostructured lipid carriers as transdermal delivery system: Part 1: Preparation, characterization and in vitro evaluation. Al Mustansiriyah J Pharm Sci. 2016;16(2):1-14. https://doi.org/10.32947/ajps.v16i2.100
[57] Wu MY, Kao IF, Fu CY, Yen SK. Effects of adding chitosan on drug entrapment efficiency and release duration for paclitaxel-loaded hydroxyapatite-gelatin composite microspheres. Pharmaceutics. 2023;15(8):2025. https://doi.org/10.3390/pharmaceutics15082025
[58] Baati T, Schembri T, Villard C, Correard F, Braguer D, Estève MA. An ultrasensitive LC-MS/MS method with liquid phase extraction to determine paclitaxel in both cell culture medium and lysate promising quantification of drug nanocarriers release in vitro. J Pharm Biomed Anal. 2015;115:300-306. https://doi.org/10.1016/j.jpba.2015.07.041
[59] Wu Z, Zou X, Yang L, Lin S, Fan J, Yang B, Sun X, Wan Q, Chen Y, Fu S. Thermosensitive hydrogel used in dual drug delivery system with paclitaxel-loaded micelles for in situ treatment of lung cancer. Colloids Surf B Biointerfaces. 2014;122:90-98. https://doi.org/10.1016/j.colsurfb.2014.06.052
[60] Al Hazzaa SA, Rajab NA. Cilnidipine Nanocrystals, Formulation and evaluation for optimization of solubility and dissolution rate. Iraqi J Pharm Sci. 2023;32(Suppl.):127-135. https://doi.org/10.31351/vol32issSuppl.pp127-135
[61] Ali SK, Al-Akkam EJ. Bilosomes as soft nanovesicular carriers for ropinirole hydrochloride: Preparation and in-vitro characterization. Iraqi J Pharm Sci. 2023;32(Suppl.):177-187. https://doi.org/10.31351/vol32issSuppl.pp177-187
[62] Salim FF, Rajab NA. Formulation and characterization of piroxicam as self-nano emulsifying drug delivery system. Iraqi J Pharm Sci. 2020;29(1):174-183. https://doi.org/10.31351/vol29iss1pp174-183
[63] Albassam NY, Kassab HJ. Diacerein loaded novasome for transdermal delivery: Prepartion, in-vitro characterization and factors affecting formulation. Iraqi J Pharm Sci. 2023;32(Suppl.):214-224. https://doi.org/10.31351/vol32issSuppl.pp214-224
[64] Jasim IK, Abd Alhammid SN, Abdulrasool AA. Synthesis and evaluation of B-cyclodextrin based nanosponges of 5-Fluorouracil by using ultrasound assisted method. Iraqi J Pharm Sci. 2020;29(2):88-98. https://doi.org/10.31351/vol29iss2pp88-98
[65] Abdullah TM, Khalid KA-K. Topical propranolol hydrochloride nanoemulsion: A promising approach drug delivery for infantile hemangiomas. Iraqi J Pharm Sci. 2023;32(Suppl.):300-315. https://doi.org/10.31351/vol32issSuppl.pp300-315
[66] Bhatta R, Chandasana H, Chhonker Y, Rathi C, Kumar D, Mitra K, Shukla P. Mucoadhesive nanoparticles for prolonged ocular delivery of natamycin: in vitro and pharmacokinetics studies. Int J Pharm. 2012;432(1-2):105-112. https://doi.org/10.1016/j.ijpharm.2012.04.060
[67] Hooda A, Nanda A, Jain M, Kumar V, Rathee P. Optimization and evaluation of gastroretentive ranitidine HCl microspheres by using design expert software. Int J Biol Macromol. 2012;51(5):691-700. https://doi.org/10.1016/j.ijbiomac.2012.07.030

Details

Primary Language

English

Subjects

Pharmacology and Pharmaceutical Sciences (Other)

Journal Section

Research Article

Authors

Mustafa Egla This is me
0009-0009-7033-283X
Iraq

Nawal Rajab ^* This is me
0000-0003-1812-2244
Iraq

Publication Date

August 3, 2026

Submission Date

March 20, 2024

Acceptance Date

June 19, 2024

Published in Issue

Year 2026 Volume: 30 Number: 4

IZ

https://izlik.org/JA97AJ43LB

Cite

RIS / Bibtex

APA

Egla, M., & Rajab, N. (2026). Screening of the most efficient variables for the development of protein nanoparticles for anticancer drug by the design of the experiment. Journal of Research in Pharmacy, 30(4), 1124-1139. https://izlik.org/JA97AJ43LB

AMA

1.Egla M, Rajab N. Screening of the most efficient variables for the development of protein nanoparticles for anticancer drug by the design of the experiment. J. Res. Pharm. 2026;30(4):1124-1139. https://izlik.org/JA97AJ43LB

Chicago

Egla, Mustafa, and Nawal Rajab. 2026. “Screening of the Most Efficient Variables for the Development of Protein Nanoparticles for Anticancer Drug by the Design of the Experiment”. Journal of Research in Pharmacy 30 (4): 1124-39. https://izlik.org/JA97AJ43LB.

EndNote

Egla M, Rajab N (August 1, 2026) Screening of the most efficient variables for the development of protein nanoparticles for anticancer drug by the design of the experiment. Journal of Research in Pharmacy 30 4 1124–1139.

IEEE

[1]M. Egla and N. Rajab, “Screening of the most efficient variables for the development of protein nanoparticles for anticancer drug by the design of the experiment”, J. Res. Pharm., vol. 30, no. 4, pp. 1124–1139, Aug. 2026, [Online]. Available: https://izlik.org/JA97AJ43LB

ISNAD

Egla, Mustafa - Rajab, Nawal. “Screening of the Most Efficient Variables for the Development of Protein Nanoparticles for Anticancer Drug by the Design of the Experiment”. Journal of Research in Pharmacy 30/4 (August 1, 2026): 1124-1139. https://izlik.org/JA97AJ43LB.

JAMA

1.Egla M, Rajab N. Screening of the most efficient variables for the development of protein nanoparticles for anticancer drug by the design of the experiment. J. Res. Pharm. 2026;30:1124–1139.

MLA

Egla, Mustafa, and Nawal Rajab. “Screening of the Most Efficient Variables for the Development of Protein Nanoparticles for Anticancer Drug by the Design of the Experiment”. Journal of Research in Pharmacy, vol. 30, no. 4, Aug. 2026, pp. 1124-39, https://izlik.org/JA97AJ43LB.

Vancouver

1.Mustafa Egla, Nawal Rajab. Screening of the most efficient variables for the development of protein nanoparticles for anticancer drug by the design of the experiment. J. Res. Pharm. [Internet]. 2026 Aug. 1;30(4):1124-39. Available from: https://izlik.org/JA97AJ43LB