Antitumor Activity of PEGylated Nanoliposomes Containing Crocin in Mice Bearing C26 Colon Carcinoma

Abstract

Crocin is a pharmacologically active component of Crocus sativus. It is an unusual water-soluble carotenoid responsible for the red color of saffron. In various studies, the anticancer effect of saffron and its constituents has been established. Polyethylene glycolated nanoliposomes with a size range up to 200 nm are suitable for encapsulation of cytotoxic drugs and can target tumors passively through the enhanced permeation and retention effect. The aim of this study was to develop a nanoliposomal formulation containing crocin with a higher therapeutic index for the treatment of cancer. Four formulations of polyethylene glycolated nanoliposomes containing 25 mg/ml crocin were prepared with hydrogenated soy phosphatidylcholine, cholesterol, and methoxy-polyethylene glycol (MW 2000)-distearoylphosphatidylcholine at different molar ratios by a solvent evaporation method plus extrusion. Then the liposomes were characterized for their size, zeta potential, crocin encapsulation, release properties, and in vitro cytotoxicity against C26 colon carcinoma cells. Based on in vitro results, the best formulation was selected for an in vivo study, and its antitumor activity was evaluated in BALB/c mice bearing C26 colon carcinoma. The IC50 of crocin itself against C26 colon carcinoma was 0.73 mM. The characterization of the best formulation was as follow: Z-average size: 127.6 ± 1.5 nm; polydispersity index: 0.087 ± 0.018; zeta potential: - 21.7 mV ± 6.7; % encapsulation: 84.62 ± 0.59; % release after 168 hours in RPMI 1640 containing 30 % FBS: 16.26 ± 0.01 %. Liposomal crocin at doses of 50 and 100 mg/kg significantly decreased tumor size and increased survival rate compared with PBS and crocin in buffer (100 mg/kg) groups. The results of this study indicated that liposomal encapsulation of crocin could increase its antitumorigenic activity. Thus, to obtain an optimal dose for use in humans, the formulation merits further investigation.

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“…Liposome preparation: Liposomes encapsulating safranal were prepared by hydration of thin lipid film followed by sonication and extrusion 15 . To put it briefly, lipid mixtures (HSPC, cholesterol and MPEG 2000 -DSPE) were dissolved in chloroform.…”

“…Safranal at different concentrations (2, 1, 0.5, 0.25 and 0.125mM) were added triplicate in each well and the cells were incubated for 72h at 37ºC. Cytotoxicity was assessed using 3-(4, 5-Dimethylthiazol-2-yl)-2, 5-diphenyl-tetrazolium bromide (MTT) assay 15 . Liposomal safranal therapy in C26 mouse model: C26 murine colorectal cells (3×10 5 ) in 50 µL PBS were inoculated S.C in the right hind flank of BALB/c mice.…”

“…Liposomal safranal therapy in C26 mouse model: C26 murine colorectal cells (3×10 5 ) in 50 µL PBS were inoculated S.C in the right hind flank of BALB/c mice. On day 10 after tumor implantation, mice were classified into different treatment groups (n=5) including liposomal safranal (50 mg/kg), safranal (50 mg/kg in Histidine buffer), isotonic PBS (200μL), Caelyx ® (15mg/kg) and doxorubicin ® (10mg/kg) 15,16 . Treatments were administered by the tail vein injection on alternate day in 5 doses for the first three groups and single dose for the other ones.…”

Safranal, a Crocus sativus L constituent suppresses the growth of K-562 cells of chronic myelogenous leukemia. In silico and in vitro study

“…A growing body of laboratory evidence indicates that saffron possesses anticancer activity against a wide spectrum of tumors, such as leukemia, gastric adenocarcinoma cancer, colon adenocarcinoma, ovarian carcinoma, hepatocellular carcinoma, rhabdomyosarcoma, lung cancer, squamous cell carcinoma and soft tissue sarcoma (Abdullaev, 2002;Abdullaev and Espinosa-Aguirre, 2004;Abdullaev and Frenkel, 1992b;Amin et al, 2011;Bathaie et al, 2013aBathaie et al, , 2013bFernández, 2006;Hoshyar et al, 2013;Molnár et al, 2000;Nair et al, 1991;Rastgoo et al, 2013). The anti-carcinogenic activity of saffron was reported in the beginning of 1990 and research on this subject has increasingly continued during the past decade.…”

“…The anti-carcinogenic activity of saffron was reported in the beginning of 1990 and research on this subject has increasingly continued during the past decade. Saffron and its main ingredients have shown antitumor and anticarcinogenic activities both in vitro and in vivo (Bathaie et al, Contents lists available at ScienceDirect 2013a, 2013bEscribano-Alonso et al, 1996;Rastgoo et al, 2013;Samarghandian et al, 2010;Tarantilis et al, 1994;Tavakkol-Afshari et al, 2008). CRC, one of the major constituents of saffron with known anticancer activity, has been found to inhibit the growth of K-562 cells of CML which express the Bcr-Abl protein tyrosine kinase (Morjani et al, 1990).…”