Lipophilic 5-fluorouracil prodrug encapsulated xylan-stearic acid conjugates nanoparticles for colon cancer therapy

doi:10.1016/j.ijbiomac.2019.01.101

International Journal of Biological Macromolecules

Volume 128, 1 May 2019, Pages 204-213

https://doi.org/10.1016/j.ijbiomac.2019.01.101 Get rights and content

Highlights

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Self-assembled nanoparticles based on Xylan-Stearic conjugates were developed.
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Self-assembled behavior was investigated using pyrene as fluorescent probe.
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The lipophilic 5-FUSA prodrug was encapsulated into the Xyl-SA NPs.
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Xyl-SA/5-FUSA NPs are biocompatible and safe for intravenous administration.
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The Xyl-SA/5-FUSA NPs induced higher cytotoxicity than free drug.

Abstract

In this study, self-assembled nanoparticles based on amphiphilic xylan-stearic acid (Xyl-SA) conjugates have been developed for the efficient delivery of 5-fluorouracil (5-FU) in cancer therapy. The self-assembled behavior of Xyl-SA conjugates in aqueous medium was investigated using pyrene as fluorescent probe. To enhance the loading efficacy of 5-FU, the lipophilic 5-fluorouracil-stearic acid (5-FUSA) prodrug was synthesized and subsequently encapsulated into the hydrophobic core of Xyl-SA NPs. The obtained Xyl-SA/5-FUSA NPs had an appropriate size (~278 nm), high drug loading of 5-FUSA (~14.6 wt%) and high physiological stability. The interaction of the Xyl-SA/5-FUSA NPs with blood components was investigated by hemolysis study. The cell cytotoxic studies demonstrated that Xyl-SA/5-FUSA NPs induced higher cytotoxicity than free drugs against the Human colorectal cancer cells (HT-29, HCT-15). These results indicate that Xyl-SA/5-FUSA NPs can serve as a promising drug delivery system for the efficient delivery of 5-FU in cancer therapy.

Introduction

Cancer is one of the serious and major health problem in the world [1,2]. Among the various cancers, colon cancer is highly prevalent and third most leading causes of death in both men and women [3]. 5-FU is one of the most prescribed chemotherapeutic agent used in colon cancer therapy, however its short half-life, poor bioavaibility, adverse sides effects and low encapsulation efficacy, limits its clinic applications [[4], [5], [6], [7]]. In order to extend the half-life or prolong circulation time in-vivo, enhance accumulation in tumor sites through enhanced permeability and retention (EPR) effect and improve the loading efficacy, various nano sized drug delivery systems including liposomes, micelles, lipid based nanoparticles and polymeric nanoparticles have been developed in recent years [8]. Usually, 5-FU shows lower encapsulation efficacy in hydrophobic core of polymeric nanoparticles due to its hydrophilic nature [9,10]. The encapsulation of 5-FU in hydrophobic core of polymeric materials could be made feasible through lipid-drug conjugate (lipophilic prodrug) approach [4]. Numerous lipids such as docosahexaenoic acid, squalenoic acid, palmitic acid and stearic acid have been explored widely in drug delivery applications because of their excellent biocompatibility and low cytotoxicity [[11], [12], [13]].

In recent years, self-assembled nanoparticles based on grafting of hydrophobic moieties to the hydrophilic back bone of polymers have received considerable interest as a potential carrier in drug delivery due to their unique core-shell architecture [1,[14], [15], [16], [17]]. The hydrophobic core of nanoparticles can be utilized as a cargo space for poorly water-soluble drugs, while the hydrophilic outer shell allows a long circulation of drug by preventing the interactions between the core and the blood components [18]. A wide range of polymers synthetic as well as natural have been used in the development of self-assembled nanoparticles [[19], [20], [21], [22], [23], [24]].

In particular, xylan is a plant cell extracted natural biopolymer and recently used in various biological applications because of its excellent physiological and biological properties [25,26]. However, due to the hydrophilic nature of xylan, it cannot form self-assembled nanoparticles in aqueous system [27]. Therefore, to get the self-assembled nanoparticles, xylan was grafted with hydrophobic moiety such as stearic acid, which could form self-assembled nanoparticles in aqueous medium [28]. In this study, lipophilic 5-FU-stearic acid (5-FUSA) prodrug was synthesized to enhance the loading efficacy of 5-FU. Towards this aim first, xylan-stearic acid conjugates nanoparticles (Xyl-SA NPs) were prepared and then lipophilic 5-FUSA prodrug was encapsulated into the hydrophobic core Xyl-SA NPs through dialysis membrane method (Scheme 1). The inner core of Xyl-SA/5-FUSA NPs formed by hydrophobic moieties (stearic acid) encapsulates the lipophilic 5-FUAC prodrug through hydrophobic interaction and outer shell formed by hydrophilic moieties (Xylan) protects the loaded drug from non-specific uptake by reticuloendothelial system (RES), resulting in an improved circulation half-life of free drug (5-FU) and minimizing its side effects on healthy cells. Moreover, due to their nano size, they can penetrate deep in the tumor tissues through fine capillaries and take up efficiently by the cancer cells thus enhanced the therapeutic efficacy of drug (5-FU) [29,30] . The obtained Xyl-SA/5-FUSA NPs were characterized and further blood compatibility and in-vitro antitumor activity of Xyl-SA/5-FUSA NPs were also investigated for their potential application in cancer therapy.

Section snippets

Materials

5-fluorouracil (≥99%), formaldehyde solution (37 wt%), dialysis membrane (MWCO 12 kDa) and 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyl tetrazolium bromide (MTT) were received from Sigma-Aldrich (Bangalore, India). Stearic acid, 4-dimethylaminopyridine (DMAP) and N, N′-dicyclohexylcarbodiimide (DCC) were obtained from Himedia (Mumbai, India). The Human colorectal cancer cells (HT-29 and HCT-15) were obtained from NCCS Pune (India). All other reagents and solvents were of analytical grade and

Synthesis and characterization of Xyl-SA conjugates and lipophilic 5-FUSA prodrug

The synthesis of Xyl-SA conjugates was confirmed by FT-IR and ¹H NMR spectroscopy. The FT-IR spectra of stearic acid exhibited the characteristics absorption bonds at 2920 cm⁻¹, 2850 cm⁻¹ and 1700 cm⁻¹ corresponding to the stretching frequencies of the single bond CH₃, CH₂ and C double bond O groups (Fig. 2a). In the xylan spectrum, the peak at 3420 cm⁻¹ is attributed to the stretching of OH group and the sharp band at 1632 cm⁻¹due to absorbed water, while the peaks between 1465 and 1043 cm⁻¹ arise from the stretching

Conclusion

The lipophilic prodrug encapsulated nano drug delivery system is a special kind of drug delivery system, which improves the loading efficacy of hydrophilic drug within the hydrophobic core of polymeric nanoperticulate. In this study, self-assembled nanoparticles based on amphiphilic xylan-stearic acid (Xyl-SA) conjugates have been developed successfully for the efficient delivery of 5-FU in cancer therapy. The loading efficacy of the hydrophilic drug (5-FU) into the hydrophobic core of Xyl-SA

Conflicts of interest

The authors declare they have no conflicts of interest.

Acknowledgments

The authors would like to thanks the MHRD New Delhi for a Ph.D. scholarship. The authors also thanks department of chemistry (IIT-Roorkee) for their convenience helps.

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Lipophilic 5-fluorouracil prodrug encapsulated xylan-stearic acid conjugates nanoparticles for colon cancer therapy

Highlights

Abstract

Introduction

Section snippets

Materials

Synthesis and characterization of Xyl-SA conjugates and lipophilic 5-FUSA prodrug

Conclusion

Conflicts of interest

Acknowledgments

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