http://informahealthcare.com/lpr ISSN: 0898-2104 (print), 1532-2394 (electronic) J Liposome Res, Early Online: 1–13 ! 2014 Informa Healthcare USA, Inc. DOI: 10.3109/08982104.2014.899369 RESEARCH ARTICLE Multifunctional liposomes for nasal delivery of the anti-Alzheimer drug tacrine hydrochloride Giuseppe Corace 1 , Cristina Angeloni 2 , Marco Malaguti 2 , Silvana Hrelia 2 , Paul C. Stein 3 , Martin Brandl 3 , Roberto Gotti 1 , and Barbara Luppi 1 1 Department of Pharmacy and Biotechnology, University of Bologna, Via San Donato, Bologna, Italy, 2 Department of Sciences for Quality of Life, University of Bologna, Corso d’Augusto, Rimini, Italy, and 3 Department of Physics, Chemistry and Pharmacy, University of Southern Denmark, Campusvej, Odense M, Denmark Abstract The purpose of this study was the development of multifunctional liposomes for nasal administration of tacrine hydrochloride. Liposomes were prepared using traditional excipients (cholesterol and phosphatidylcholine), partly enriched with a-tocopherol and/or Omega3 fatty acids. This approach was chosen in order to obtain at the same time two positive results: an enhanced drug permeation through nasal mucosa and a concomitant neuroprotective effect. Several liposome formulations were prepared using the Reverse Phase Evaporation technique followed by membrane filter extrusion. In particular, liposome capacity to enhance drug permeation was evaluated by means of membrane permeation and cellular uptake studies. Furthermore, liposome effect on neuronal viability and intracellular ROS production was evaluated as well as their cytoprotective effect against oxidative stress. All liposome formulations showed a mean diameter in the range of 175 nm to 219 nm with polydispersity index lower than 0.22, a lightly negative zeta potential and excellent encapsulation efficiency. Moreover, along with good mucoadhesive properties, multifunctional liposomes showed a markedly increase in tacrine permeability, which can be related to liposome fusion with cellular membrane, a hypothesis, which was also supported by cellular uptake studies. Finally, the addition of a-tocopherol without Omega3 fatty acids, was found to increase the neuroprotective activity and antioxidant properties of liposomes. Keywords Antioxidant activity, enhanced permeation, nasal administration, neuronal uptake History Received 13 December 2013 Revised 21 February 2014 Accepted 25 February 2014 Published online 7 May 2014 Introduction At present, there are no therapeutic interventions able to stop the progression of Alzheimer’s disease (AD) or to treat progressive degeneration of the acetylcholine neurons of the brain, which leads to the appearance of cognitive symptoms of the disease. Treatments are therefore aimed at slowing down the worsening course, maintaining, at least temporarily, the cognitive and behavioral symptoms, thus ensuring patients to have good or decent quality of life. One strategy to combat this disease is to increase cholinergic transmission; therefore acetylcholinesterase inhibitors have been developed including tacrine, rivastigmine and donepezil (Giacobini, 1993; Polinsky, 1998). Although the aetiology is not still under- stood, it appears that oxidative stress (OS) plays a central role in the pathogenesis and progression of the AD (Guglielmotto et al., 2010) and then it would be desirable to integrate traditional therapeutic strategy, based on the recovery of the acetylcholine deficit, with an antioxidant treatment (Minarini et al., 2012). In fact, epidemiological investigations have shown an improvement in cognitive activity and a slowing of disease progression following the administration of antioxi- dants (Masaki et al., 2000). In particular, a-tocopherol (Toc) seems to be a suitable choice for AD treatment when performed with the anti-Alzheimer drug Tacrine (THA), as Toc co-administration has been shown to reduce the hepato- toxic effects of THA (Dogterom et al., 1988). Another important factor for the brain health is its content in docosahexaenoic acid (DHA), an Omega3 fatty acid (V3) biosynthesized also starting from a-linolenic acid. When the brain is rich in DHA, the characteristic symptoms of the disease decrease due to the lower production of senile plaques (Yazawa, 1996). This fact suggests that co-administration of anti-Alzheimer drugs and V3 can be a valid approach promoting increased beneficial effect towards the disease. THA, one of the drugs approved by the FDA for the treatment of AD (Davis & Powchik, 1995; Summers, 2006), is present in the market only as oral formulation. However, THA administration is associated with low bioavailability, due to the extensive hepatic first-pass effect and the rapid clearance from the systemic circulation (Hartvig et al., 1990). THA administration is also associated with a reversible Address for correspondence: Barbara Luppi, Department of Pharmacy and Biotechnology, University of Bologna, Via San Donato 19/2, 40127, Bologna, Italy. Tel: +39 0512095198. Fax: +39 0512095199. E-mail: barbara.luppi@unibo.it Journal of Liposome Research Downloaded from informahealthcare.com by D S Diffusioni Scientifiche - Unical on 05/16/14 For personal use only.