Nanoemulsions of cancer chemopreventive agent benzyl isothiocyanate display enhanced solubility, dissolution, and permeability.

Benzyl isothiocyanate (BITC), a compound found in cruciferous vegetables, is an effective chemopreventive agent. The objective of this study was to develop nanoemulsion formulations for the oral delivery of BITC. Optimized oil-in-water BITC nanoemulsions were prepared by a spontaneous self-nanoemulsification method and a homogenization-sonication method. Both nanoemulsions entrapped high amounts of BITC (15-17 mg/mL), with low polydispersity and good colloidal stability. The BITC nanoemulsions showed enhanced solubility and dissolution compared to pure BITC. These formulations markedly increased the apical to basolateral transport of BITC in Caco-2 cell monolayers. The apparent permeability values were 3.6 × 10(-6) cm/s for pure BITC and (1.1-1.3) × 10(-5) cm/s for BITC nanoemulsions. The nanoemulsions were easily taken up by human cancer cells A549 and SKOV-3 and inhibited tumor growth in vitro. This work shows for the first time that BITC can be formulated into nanoemulsions and may show promise in enhancing absorption and bioavailability.

[1]  A. Chow,et al.  Self-nanoemulsifying drug delivery system (SNEDDS) for oral delivery of Zedoary essential oil: formulation and bioavailability studies. , 2010, International journal of pharmaceutics.

[2]  D. Waitzberg,et al.  New parenteral lipid emulsions for clinical use. , 2006, JPEN. Journal of parenteral and enteral nutrition.

[3]  A. Misra,et al.  Formulation and Characterization of Nanoemulsion-Based Drug Delivery System of Risperidone , 2009, Drug development and industrial pharmacy.

[4]  L. Walker,et al.  Flax seed oil and flax seed meal reduce the formation of aberrant crypt foci (ACF) in azoxymethane-induced colon cancer in Fisher 344 male rats. , 2007, Food and chemical toxicology : an international journal published for the British Industrial Biological Research Association.

[5]  P. Artursson,et al.  Correlation between oral drug absorption in humans and apparent drug permeability coefficients in human intestinal epithelial (Caco-2) cells. , 1991, Biochemical and biophysical research communications.

[6]  Steven W. Booth,et al.  An investigation into the mechanisms of self-emulsification using particle size analysis and low frequency dielectric spectroscopy , 1995 .

[7]  S. Ganta,et al.  Coadministration of Paclitaxel and curcumin in nanoemulsion formulations to overcome multidrug resistance in tumor cells. , 2009, Molecular pharmaceutics.

[8]  P Augustijns,et al.  Effect of simulated intestinal fluid on drug permeability estimation across Caco-2 monolayers. , 2004, International journal of pharmaceutics.

[9]  F. Shakeel,et al.  Investigation of true nanoemulsions for transdermal potential of indomethacin: characterization, rheological characteristics, and ex vivo skin permeation studies , 2009, Journal of drug targeting.

[10]  M. Lawrence,et al.  Microemulsion-based media as novel drug delivery systems , 2000 .

[11]  S. Baboota,et al.  Formulation development and optimization using nanoemulsion technique: A technical note , 2007, AAPS PharmSciTech.

[12]  F. Chung,et al.  Isothiocyanates as cancer chemopreventive agents: their biological activities and metabolism in rodents and humans. , 2002, Current drug metabolism.

[13]  K. Pathak,et al.  Therapeutic applications of nanoemulsion based drug delivery systems: a review of patents in last two decades. , 2011, Recent patents on drug delivery & formulation.

[14]  M. Rizwan,et al.  Nanoemulsion Components Screening and Selection: a Technical Note , 2009, AAPS PharmSciTech.

[15]  Makoto Kataoka,et al.  In Vitro System to Evaluate Oral Absorption of Poorly Water-Soluble Drugs: Simultaneous Analysis on Dissolution and Permeation of Drugs , 2003, Pharmaceutical Research.

[16]  Hetal Thakkar,et al.  Formulation and characterization of lipid-based drug delivery system of raloxifene-microemulsion and self-microemulsifying drug delivery system , 2011, Journal of pharmacy & bioallied sciences.

[17]  L. Wattenberg.  Inhibition of carcinogenic effects of polycyclic hydrocarbons by benzyl isothiocyanate and related compounds. , 1977, Journal of the National Cancer Institute.

[18]  Arik Dahan,et al.  Rationalizing the selection of oral lipid based drug delivery systems by an in vitro dynamic lipolysis model for improved oral bioavailability of poorly water soluble drugs. , 2008, Journal of controlled release : official journal of the Controlled Release Society.

[19]  Hailong Yu,et al.  Encapsulation of epigallocatechin-3-gallate (EGCG) using oil-in-water (O/W) submicrometer emulsions stabilized by ι-carrageenan and β-lactoglobulin. , 2010, Journal of agricultural and food chemistry.

[20]  E. Mantzioris,et al.  The effect on human tumor necrosis factor alpha and interleukin 1 beta production of diets enriched in n-3 fatty acids from vegetable oil or fish oil. , 1996, The American journal of clinical nutrition.

[21]  S. Majumdar,et al.  Injectable Lipid Emulsions—Advancements, Opportunities and Challenges , 2010, AAPS PharmSciTech.

[22]  Tushar K. Vyas,et al.  Improved oral bioavailability and brain transport of Saquinavir upon administration in novel nanoemulsion formulations. , 2008, International journal of pharmaceutics.

[23]  David Julian McClements,et al.  Formation of flavor oil microemulsions, nanoemulsions and emulsions: influence of composition and preparation method. , 2011, Journal of agricultural and food chemistry.

[24]  Mushir M. Ali,et al.  Nanocarrier for the enhanced bioavailability of a cardiovascular agent: in vitro, pharmacodynamic, pharmacokinetic and stability assessment. , 2011, International journal of pharmaceutics.

[25]  S. Batra,et al.  Benzyl isothiocyanate-mediated generation of reactive oxygen species causes cell cycle arrest and induces apoptosis via activation of MAPK in human pancreatic cancer cells. , 2009, Carcinogenesis.

[26]  Yong-Hae Han,et al.  Current industrial practices of assessing permeability and P-glycoprotein interaction , 2006, The AAPS Journal.

[27]  C. Solans,et al.  Formation of Nano-emulsions by Low-Energy Emulsification Methods at Constant Temperature , 2001 .

[28]  S. Batra,et al.  Benzyl Isothiocyanate–Mediated Inhibition of Histone Deacetylase Leads to NF-κB Turnoff in Human Pancreatic Carcinoma Cells , 2010, Molecular Cancer Therapeutics.

[29]  S. Hecht,et al.  High-performance liquid chromatography-based determination of total isothiocyanate levels in human plasma: application to studies with 2-phenethyl isothiocyanate. , 2001, Analytical biochemistry.

[30]  Tushar K. Vyas,et al.  Cytotoxicity and apoptosis enhancement in brain tumor cells upon coadministration of paclitaxel and ceramide in nanoemulsion formulations. , 2008, Journal of pharmaceutical sciences.

[31]  C. Porter,et al.  LIPID-BASED FORMULATIONS FOR ORAL ADMINISTRATION: OPPORTUNITIES FOR BIOAVAILABILITY ENHANCEMENT AND LIPOPROTEIN TARGETING OF LIPOPHILIC DRUGS , 2001, Journal of receptor and signal transduction research.

[32]  J. Crison,et al.  A Theoretical Basis for a Biopharmaceutic Drug Classification: The Correlation of in Vitro Drug Product Dissolution and in Vivo Bioavailability , 1995, Pharmaceutical Research.

[33]  B. Koletzko,et al.  Parenteral lipid emulsions in paediatrics , 2006, Current opinion in clinical nutrition and metabolic care.

[34]  Koji Takahashi,et al.  Microemulsion formulation for enhanced absorption of poorly soluble drugs. I. Prescription design. , 2002, Journal of controlled release : official journal of the Controlled Release Society.

[35]  D. Lairon,et al.  Digestion and absorption of 2 fat emulsions with different droplet sizes in the human digestive tract. , 1999, The American journal of clinical nutrition.

[36]  L. Kolonel,et al.  Vegetable consumption and lung cancer risk: a population-based case-control study in Hawaii. , 1989, Journal of the National Cancer Institute.

[37]  P. van Bladeren,et al.  Association between consumption of cruciferous vegetables and condiments and excretion in urine of isothiocyanate mercapturic acids. , 2006, Journal of agricultural and food chemistry.

[38]  Thierry F. Vandamme,et al.  Nano-emulsions and Micro-emulsions: Clarifications of the Critical Differences , 2011, Pharmaceutical Research.

[39]  Susan O'Brien,et al.  NCCN Task Force Report: Oral chemotherapy. , 2008, Journal of the National Comprehensive Cancer Network : JNCCN.

[40]  D. Scudiero,et al.  New colorimetric cytotoxicity assay for anticancer-drug screening. , 1990, Journal of the National Cancer Institute.

[41]  D. Mcclements,et al.  Food-Grade Nanoemulsions: Formulation, Fabrication, Properties, Performance, Biological Fate, and Potential Toxicity , 2011, Critical reviews in food science and nutrition.

[42]  D. Horne,et al.  Structuring food emulsions in the gastrointestinal tract to modify lipid digestion. , 2009, Progress in lipid research.

[43]  C. Dwivedi,et al.  Chemopreventive Effects of Dietary Flaxseed on Colon Tumor Development , 2006, Nutrition and cancer.

[44]  K. Luthman,et al.  Caco-2 monolayers in experimental and theoretical predictions of drug transport , 1996 .

[45]  The role of STAT-3 in the induction of apoptosis in pancreatic cancer cells by benzyl isothiocyanate. , 2009, Journal of the National Cancer Institute.

[46]  J. Panyam,et al.  Injectable sustained release microparticles of curcumin: a new concept for cancer chemoprevention. , 2010, Cancer research.

[47]  S. Hecht,et al.  Chemoprevention by isothiocyanates , 1995, Journal of cellular biochemistry. Supplement.

[48]  Alex Avdeef,et al.  In vitro trans-monolayer permeability calculations: often forgotten assumptions. , 2003, Drug discovery today.

[49]  Y. Masaoka,et al.  Effect of food intake on the oral absorption of poorly water-soluble drugs: in vitro assessment of drug dissolution and permeation assay system. , 2006, Journal of pharmaceutical sciences.

[50]  Kristina Luthman,et al.  Caco-2 monolayers in experimental and theoretical predictions of drug transport1PII of original article: S0169-409X(96)00415-2. The article was originally published in Advanced Drug Delivery Reviews 22 (1996) 67–84.1 , 2001 .

[51]  U Norinder,et al.  Experimental and computational screening models for the prediction of intestinal drug absorption. , 2001, Journal of medicinal chemistry.

[52]  S. B. Tiwari,et al.  Improved oral delivery of paclitaxel following administration in nanoemulsion formulations. , 2006, Journal of nanoscience and nanotechnology.

[53]  M. Coelho,et al.  Epigallocatechin gallate-loaded polysaccharide nanoparticles for prostate cancer chemoprevention. , 2011, Nanomedicine.

[54]  L. Walker,et al.  Red palm oil suppresses the formation of azoxymethane (AOM) induced aberrant crypt foci (ACF) in Fisher 344 male rats. , 2006, Food and chemical toxicology : an international journal published for the British Industrial Biological Research Association.

[55]  P. Sanguansri,et al.  Impact of oil type on nanoemulsion formation and Ostwald ripening stability. , 2008, Langmuir : the ACS journal of surfaces and colloids.

[56]  D. Rose,et al.  Omega-3 fatty acids as cancer chemopreventive agents. , 1999, Pharmacology & therapeutics.

[57]  C. Dwivedi,et al.  Chemopreventive effects of dietary flaxseed on azoxymethane-induced colon tumor development in rats , 2005 .

[58]  D. Lorenz,et al.  Studies on the metabolism and excretion of benzyl isothiocyanate in man. , 1988, Xenobiotica; the fate of foreign compounds in biological systems.

[59]  P. Artursson,et al.  Determination of drug permeability and prediction of drug absorption in Caco-2 monolayers , 2007, Nature Protocols.

[60]  R. Goldbohm,et al.  Epidemiological studies on brassica vegetables and cancer risk. , 1996, Cancer epidemiology, biomarkers & prevention : a publication of the American Association for Cancer Research, cosponsored by the American Society of Preventive Oncology.