International Journal of Pharmaceutics 454 (2013) 649–652 Contents lists available at ScienceDirect International Journal of Pharmaceutics j o ur nal ho me page: www.elsevier.com/locate/ijpharm Note on the formulation of thermosensitive and mucoadhesive vaginal hydrogels containing the miniCD4 M48U1 as anti-HIV-1 microbicide Kawthar Bouchemal a,∗ , Justyna Frelichowska a , Loïc Martin b , Vanessa Lievin-Le Moal c , Roger Le Grand d , Nathalie Dereuddre-Bosquet d , Madeleine Djabourov e , Armelle Aka-Any-Grah a,f , Armand Koffi f , Gilles Ponchel a a Université Paris-Sud, Institut Galien Paris Sud, UMR CNRS 8612, Faculté de Pharmacie, 5, rue J.B. Clément, 92296 Châtenay-Malabry cedex, France b CEA, iBiTecS, Service d’Ingénierie Moléculaire des Protéines (SIMOPRO), 91191 Gif sur Yvette, France c Université Paris-Sud, UMR CNRS 8076 “Biomolécules: Conception, Isolement, Synthèse”, Groupe Chimiothérapie Antiparasitaire, Faculté de Pharmacie, 5, rue J.B. Clément, 92296 Châtenay-Malabry cedex, France d CEA, iMETI, Division of Immuno-Virology, Fontenay-aux Roses, France e Laboratoire de Physique Thermique, ESPCI-Paris Tech, 10 rue Vauquelin, 75231 Paris cedex 05, France f Laboratoire de Pharmacologie et de Pharmacocinétique, UFR des sciences pharmaceutiques et Biologiques d’Abidjan, 01 BP V 34 Abidjan 01, Télex 26138, Cote d’Ivoire a r t i c l e i n f o Article history: Received 12 January 2013 Received in revised form 25 February 2013 Accepted 26 February 2013 Available online 7 March 2013 Keywords: Pluronic ® Anti-HIV-1 microbicides M48U1 Toxicological evaluation Release profiles a b s t r a c t The miniCD4 M48U1 was formulated into thermosensitive and mucoadhesive pluronic ® hydrogels as anti-HIV-1 microbicide. The release kinetics of M48U1 from F127/HPMC (20/1 wt%) and F127/F68/HPMC (22.5/2.5/1 wt%) studied during 24 h by using Franz diffusion cells showed that HEC hydrogel (1.5 wt%) used as control released 93% of the peptide, while about 25% of M48U1 remained in pluronic ® hydrogels. The formulation of M48U1 in pluronic ® hydrogels ensures a local delivery because no diffusion of the peptide was detected through vaginal Cynomolgus macaque mucosa using Ussing chamber. Finally, tox- icological studies showed no significant difference in the HeLa cell viability of the pluronic ® hydrogels in comparison with HEC and phosphate buffer saline. © 2013 Elsevier B.V. All rights reserved. At present, the development of potent anti-HIV microbicides self-administered by women before intercourse still represents important strategy for HIV prevention. Encouraging results were obtained from the CAPRISA-004 phase IIb clinical trial of 1% teno- fovir formulated in hydroxyetlycellulose (HEC) hydrogel. Tenofovir, which is a nucleotide reverse transcriptase inhibitor, reduced HIV acquisition by an estimated 39% overall and by 54% in women with hydrogel adherence (Abdool Karim et al., 2010). However, more recently, trial of a once-daily dosing regimen with tenofovir gel (VOICE) found that the gel was not effective among study partici- pants. In this context, the group of Martin et al. has developed a novel miniCD4 as a potential HIV-1 microbicide that tar- get initial step of viral attachment to CD4 to block the viral entry (Martin et al., 2003; Van Herrewege et al., 2008; Stricher ∗ Corresponding author. Tel.: +33 01 46 83 55 81; fax: +33 01 46 61 93 34. E-mail address: kawthar.bouchemal@u-psud.fr (K. Bouchemal). et al., 2008). This peptide named M48U1 (Fig. 1) containing a p- (cyclohexylmethyloxy)phenylalanine residue at position 23 was synthesized at Pepscan Presto Inc. (Lelystad, The Netherlands) by solid phase peptide synthesis and purified after refolding by reverse-phase high performance liquid chromatography as described elsewhere (Martin et al., 2011). Thanks to its small size and the presence of a protusion stabilizing the Phe43-cavity (an additional cyclohexyl moiety), M48U1 binds to gp120 with a higher affinity than the native CD4. The objectives of the present work are: • To formulate M48U1 into thermosensitive and mucoadhesive hydrogels. It is expected that the hydrogel-containing M48U1 acts as a barrier against virus diffusion. • To evaluate the toxicity of hydrogels toward HeLa cells derived from cervical cancer cells. • To study the release profiles of M48U1 from the hydrogels. • When a local application is considered we have to check that no drug will pass through the vaginal mucosa. The permeation 0378-5173/$ – see front matter © 2013 Elsevier B.V. All rights reserved. http://dx.doi.org/10.1016/j.ijpharm.2013.02.055