Research paper Nanosuspension for the delivery of a poorly soluble anti-cancer kinase inhibitor Fabienne Danhier a , Bernard Ucakar a , Marie-Lyse Vanderhaegen a , Marcus E. Brewster b , Tina Arien b , Véronique Préat a,⇑ a Université Catholique de Louvain, Louvain Drug Research Institute, Pharmaceutics and Drug Delivery, Brussels, Belgium b Johnson and Johnson, Pharmaceutical Research and Development, Division of Janssen Pharmaceutica, Beerse, Belgium article info Article history: Received 5 February 2014 Accepted in revised form 8 May 2014 Available online xxxx Keywords: Nanosuspension Micelles Nanoparticles Tumor targeting Kinase inhibitor MTKi-327 Anti-cancer drug abstract We hypothesized that nanosuspensions could be promising for the delivery of the poorly water soluble anti-cancer multi-targeted kinase inhibitor, MTKi-327. Hence, the aims of this work were (i) to evaluate the MTKi-327 nanosuspension for parenteral and oral administrations and (ii) to compare this nanosus- pension with other nanocarriers in terms of anti-cancer efficacy and pharmacokinetics. Therefore, four formulations of MTKi-327 were studied: (i) PEGylated PLGA-based nanoparticles, (ii) self-assembling PEG 750 -p-(CL-co-TMC) polymeric micelles, (iii) nanosuspensions of MTKi-327; and (iv) Captisol solution (pH = 3.5). All the nano-formulations presented a size below 200 nm. Injections of the highest possible dose of the three nano-formulations did not induce any side effects in mice. In contrast, the maximum tolerated dose of the control Captisol solution was 20-fold lower than its highest possible dose. The high- est regrowth delay of A-431-tumor-bearing nude mice was obtained with MTKi-327 nanosuspension, administered intravenously, at a dose of 650 mg/kg. After intravenous and oral administration, the AUC 0–1 of MTKi-327 nanosuspension was 2.4-fold greater than that of the Captisol solution. Nanosus- pension may be considered as an effective anti-cancer MTKi-327 delivery method due to (i) the higher MTKi-327 maximum tolerated dose, (ii) the possible intravenous injection of MTKi-327, (iii) its ability to enhance the administered dose and (iv) its higher efficacy. Ó 2014 Elsevier B.V. All rights reserved. 1. Introduction Poor water solubility of many anti-cancer drugs or drug candidates constitutes a major obstacle to their development and their clinical application. To overcome this problem, several strategies have been developed such as the use of cyclodextrins, salt formation of ionizable drugs and the use of co-solvents or surfactants (which are often toxic; e.g. Cremophor Ò EL) [1]. Alterna- tively, numerous nanosystems have been investigated for the colloidal dispersion of poorly water soluble drugs, including lipo- somes, nanoparticles, polymeric micelles, dendrimers, etc. These nanocarriers have been largely described for their ability to deliver the drug specifically to tumor, decreasing systemic side-effects. Some nanoformulations have been clinically approved such as Abraxane Ò and Doxil Ò [2]. Because of the fenestrations present in tumoral endothelium, these nanocarriers can enter the interstit- ium, be entrapped in the tumor and remain inside the tumoral tis- sue because of the poor lymphatic drainage. This ‘‘passive’’ targeting is based on the enhanced permeability and retention (EPR) effect, discovered by Matsumura and Maeda [3]. Recently, various nanosizing strategies have emerged. Nanosus- pensions (also called nanocrystals) are nanoscopic crystals of the compound with a size below 1 lm [1]. Nanosuspensions present several advantages: (i) Nanosuspensions allow the colloidal dispersion of poorly water soluble drugs [4]. (ii) These particles possess a very high drug loading (pure drug coated with surfactant http://dx.doi.org/10.1016/j.ejpb.2014.05.014 0939-6411/Ó 2014 Elsevier B.V. All rights reserved. Abbreviations: AUC, area under the curve; CL, caprolactone; DMEM, Dubelcco’s modified Eagle’s medium; EDTA, ethylenediaminetetraacetic acid; EGFR, epidermal growth factor receptor; EMA, European Medicine Agency; EPR, enhanced perme- ability and retention effect; FDA, Food and Drug Administration; HPLC, high performance liquid chromatography; IV, intravenous; LOQ, limit of quantification; MTD, maximum tolerated dose; NMR, nuclear magnetic resonance; PCL, poly- caprolactone; PDI, polydispersity index; PEG, polyethylene glycol; PK, pharmaco- kinetic; PLGA, poly(lactide-co-glycolide); PTX, paclitaxel; PVDF, polyvinylidene fluoride; RES, reticulo-endothelial system; SEC, size exclusion chromatography; SEM, standard error of the mean; TMC, trimethylene carbonate; VEGFR, vascular endothelial growth factor receptor. ⇑ Corresponding author. Université Catholique de Louvain, Louvain Drug Research Institute, Pharmaceutics and Drug Delivery, Avenue Mounier B1.73.12, B-1200 Brussels, Belgium. Tel.: +32 2 7647320; fax: +32 2 7647398. E-mail address: veronique.preat@uclouvain.be (V. Préat). European Journal of Pharmaceutics and Biopharmaceutics xxx (2014) xxx–xxx Contents lists available at ScienceDirect European Journal of Pharmaceutics and Biopharmaceutics journal homepage: www.elsevier.com/locate/ejpb Please cite this article in press as: F. Danhier et al., Nanosuspension for the delivery of a poorly soluble anti-cancer kinase inhibitor, Eur. J. Pharm. Bio- pharm. (2014), http://dx.doi.org/10.1016/j.ejpb.2014.05.014