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J Pharm Pharmaceut Sci ualberta ~csps ; 7 2 ; : 92-185, 2004 Purpose. To exploit the solubilizing potential of hydroxypropylcellulose-g-poly ethyleneglycol ; cetyl ether HPC-g-PEG-C16 ; polymeric micelles towards poorly-water soluble drugs in order to improve their oral bioavailability. Methods. Hydrophobically-modified HPC-g-PEG-C16 graft copolymer was synthesized by attaching hydrophobic hexadecyl residues to the hydrophilic HPC backbone via short PEG linkers. Cyclosporin A CsA ; , a highly effective immunosuppressant, was selected as model drug. Its water solubility is as low as 23g mL. CsA-loaded HPC-g-PEG-C16 micelles were prepared by a dialysis procedure using initial CsA loadings of 2.5-40% w w ; . Incorporated CsA was assayed by HPLC at 210 nm. The cytotoxicity of HPC, PEG-C16 and HPC-g-PEGC16 towards the human colon adenocarcinoma, Caco-2, cells was evaluated using the MTT colorimetric assay. Transport of free CsA or micelles-loaded CsA across Caco-2 monolayers was evaluated at predetermined time points. Pluronic P85 polymer was added as P-glycoprotein inhibitor. Results. At 25% w w ; of initially added CsA, HPC-g-PEG-C16 micelles showed 22% entrapment efficiency for CsA, compared to 4% in the case of free HPC polymer. After 24 h, free HPC showed no cytotoxicity while free PEG-C16 inhibited cell growth even at low concentrations. Most importantly, the study revealed that HPCg-PEG-C16 exhibited no significant cytotoxicity at a concentration as high as 10 g Following 240 min of transport, the apical to basal permeability of CsA across caco-2 cells increased by 3 times when loaded in HPC-g-PEG-C16, compared to free CsA. Conclusions. HPC-g-PEG-C16 polymeric micelles increased the aqueous solubility of the lipophilic drug CsA. Their very low cytotoxicity and high permeability represent promising characteristics for the development of a novel polymeric drug carrier for the oral delivery of poorly-water soluble drugs. NSERC and Rx&D are acknowledged for financial support. This work has been presented at the 2003 AAPS annual meeting on October 30 and received the 2003 Award of the "Pharmaceutics and Drug Delivery" section. 60 TEXTURE PROFILE ANALYSIS AS A MEAN TO ASSESS THE MECHANICAL PROPERTIES OF THERMOREVERSIBLE MUCOADHESIVE HYDROGELS Noha Gouda, Patrice Hildgen; Facult de Pharmacie, Universit de Montral, Montral, Qubec, Canada Purpose. Based on the assessment of mechanical properties, the purpose of the current study is to select the optimum thermoreversible mucoadhesive hydrogel formulation, aimed to overcome the drawbacks of conventional nasal dosage forms such as poor residence time, leakage or difficulty of application. Methods. Aqueous solutions, of different concentrations of thermoreversible polymers poloxamer407 P407 ; and poloxamer188 P188 ; with or without the mucoadhesive polymers carbopol 934P CP934P ; or polycarbophil PC ; , were prepared. Their gelation temperatures were determined using the inverting test method. Kept at 34C0.5 for 30 minutes prior testing, three replicates from each free flowing liquid formulation were allowed to form gels. Texture profile analysis TPA ; was then performed using the texture analyser TA-XT2i Texture Technologies Corp. NY ; where 2cm diameter cylindrical acrylic probe was twice compressed into each formulation at 34C to 5.0mm depth, at pre-test speed of 5.0mm s, test and post-test speed of 1.0mm s, and recovery period of 15s between the 2 compression cycles. From the resultant forcetime curve, values of hardness, adhesiveness and cohesiveness were determined. Results were analysed using artificial neural networks. Results. Formulation with optimum mechanical properties appeared to be that containing P407 P188 PC of concentrations 15.0 25.0 0.1 %w w ; where the values of hardness, adhesiveness, and cohesiveness were 2.03 N, 7.20 Nmm, 1.06 Nmm respectively. These properties were negligible for formulations not converted into gels at body temperature. PC proved to play a considerable role in cohesiveness relative importance 83%, R2 0.946 and MSE 0, 001 ; .Cohesiveness, when weak, affects the adhesiveness. Conclusion. TPA, a simple and rapid procedure, is valuable in the scanning of the mechanical characteristics of different thermoreversible hydrogel formulations to obtain the suitable candidate for mucosal application. 61 CHARACTERIZATION OF RECOMBINANT AMINOPEPTIDASE P Yvan Tran, Giuseppe Molinaro, Albert Adam; Facult de pharmacie, Universit de Montral, succursale Centreville, Montral, Qubec, Canada Introduction To better understand the role of the metallopeptidase X-prolyl aminopeptidase APP ; in angiotensin I-converting enzyme inhibitors ACEi ; side effects, our laboratory used recombinant technology to produce human APP enzyme. The purpose of this study is to characterize the catalytic properties of this recombinant enzyme using different experimental conditions, and to compare these values to known data published elsewhere. Materials & methods Recombinant metallopeptidase catalytic activities and characteristics were assessed using various methods, namely time-point spectrofluorometry and HPLC techniques. For this, synthetic, natural and novel internally quenched fluorescent substrates were studied, and their catalytic properties kcat, Km ; determined. Optimal pH ranges for recombinant APP activity, as well as metal ion requirements were assessed. The inhibition profile of the recombinant enzyme was studied using the newly developed fluorescent substrate IC50, Ki ; . Results Optimal pH conditions determined for the recombinant enzyme closely matched those of an enzyme preparation purified from animal tissues rat, bovine & porcine ; previously published. Affinity constants of the recombinant form for its physiological. Isordil 10 mg tablet contains 10 mg isosorbide dinitrate.
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