315 PHARMACOKINETICS, PHARMACOLOGICAL STABILITY AND SAFETY OF MMC ADMINISTERED WITH A NEW HYPERTHERMIA DEVICE Paolo Gontero*, Paola Milla, Nicoletta Serra, Chiara Fiorito, Francesco Cattel, Luigi Cattel, Alessandro Tizzani, Torino, Italy INTRODUCTION AND OBJECTIVES: Hyperthermic administration of MMC with the Synergo has proved effective with acceptable tolerability profile in high risk non muscle invasive bladder cancer. The major drawback is the high costs of both the device and disposable catheters. We report data on drug stability and safety of Unithermia, a new hyperthermia device for intravesical administration of MMC. METHODS: Ten patients with a BCG recurrent intermediate risk NMIBC were enrolled in the phase I part of the study and received 6 weekly instillations of MMC Unithermia. 20 mg MMC diluted in 50 ml were adminis- tered via the Unithermia device for 45 minutes at a temperature of 42°C. A fresh drug solution was replaced at 23 minutes of every cycle. Pharmacolog- ical stability was tested by HPLC analysis of residual MMC on the drug solutions retrieved at mid and end of each instillation. Blood samples were taken at 0, 12, 23, 34 and 45 minutes time intervals for the evaluation of plasmatic pharmacokinetic of MMC. Adverse events were graded according to the CTCAEv6. Cystoscopy was planned every 3 months for the first year. RESULTS: MMC stability was evaluated on 39 cycles: the median recovery of normal MMC was 66% and 99% at 23 and 45 minutes respec- tively. Cmax plasmatic levels of MMC remained low within each time point of a single instillation and during the whole induction course. Urinary side effects did not go beyond grade 2, 1 patient developed grade 3 systemic skin reaction and 3 patients discontinued the treatment at the 4th, 5th and 5th instillation respectively. Necrotic area on the posterior bladder wall was observed in 1/10 at cystoscopy. CONCLUSIONS: The analysis of drug solutions retrieved at mid and end of instillation demonstrates that MMC stability was not affected by Unith- ermia administration, with the main drug absorption occurring during the first half of the instillation. Plasmatic MMC levels remained negligible. Discontinu- ation rates were not related to severe toxicity. Phase II studies will be planned to test for activity of the treatment. Source of Funding: None 316 SUCCESSFUL INTRAVESICAL THERAPY FOR BLADDER CANCER UTILIZING PACLITAXEL CONTAINING NANOPARTICLES Mary Samplaski*, Armine Smith, William Larchian, Vinod Labhasetwar, Warren Heston, Cleveland, OH INTRODUCTION AND OBJECTIVES: About 70% of bladder cancers are superficial and amenable to intravesical therapy. Nanopar- ticles (NPs) provide a vehicle for localized drug delivery. They achieve rapid intracellular penetration and provide slow, regional release of encapsulated drug. The transferrin receptor (TfR) is overexpressed in bladder cancer cells. Paclitaxel (Tx) has activity against human bladder cancer. To optimize NP preparation, we evaluated Tf and poly-L-lysine (PLL) conjugated NP penetration. We then tested Tf conjugated NPs for Tx delivery to bladder cancer cells in a murine model. METHODS: After bladder pretreatment with PLL, orthotopic MBT-2 bladder tumors were induced in C3H/HeJ female mice. Tumors were con- firmed sonographically at 14 days. Coumarin-conjugated NPs were instilled intravesically in several groups: with & without PLL pretreatment, conjugated & unconjugated to PLL, and conjugated & unconjugated to Tf. Confocal microscopy of bladders was done after 1.5 hours of incubation. All subsequent experiments used Tf-conjugated NPs without PLL pretreatment. Cremophor (solvent for Tx) & Tx, Cremophor & saline, saline, and Tx loaded NPs (8 mg/kg) were instilled intravesically for 1.5 hours. Ultrasound was performed starting on post-instillation day (PID) 5. Bladders were examined on PID 15. RESULTS: Tf conjugation led to enhanced NP accumulation in the tumors. PLL conjugation and PLL pre-wash enhanced overall penetration but negated the selective tumor binding. By PID 5 the Cremophor, Cremophor & saline, and saline groups had developed enlarging bladder tumors  1mm. On PID 5 and 7, tumors in the Tx-NP group were comparatively smaller, 0.682mm and 0.878mm, respectively. By PID 9 tumors in the Tx-NP group were also enlarging (Fig 1). Histolology confirmed invasive tumors in all groups on PID 15. CONCLUSIONS: Tf-NP conjugation allows for selective uptake of the NP by cancer cells. A single dose of NP encapsulated Tx suppresses murine superficial bladder tumor growth for 7 days. Further studies will focus on repeated dosing of Tx-NPs and increasing the dose of Tx. Source of Funding: None 317 EFFECT OF ATORVASTATIN ON THE BIOLOGY OF BLADDER CANCER CELLS TREATED WITH BACILLUS CALMETTE-GUERIN Tullika Garg*, Guangjian Zhang, Fanghong Chen, Yanli Cao, William See, Milwaukee, WI INTRODUCTION AND OBJECTIVES: The commonly employed class of cholesterol lowering agents, statins, functions by inhibiting the HMG- CoA reductase enzyme which is critical to cholesterol biosynthesis. Early clinical reports suggest that BCG efficacy may be decreased in patients taking statins. This study examined the effect of BCG on enzymes in the cholesterol synthesis pathway, the impact of statins on the BCG induced signaling pathways and gene expression, as well as the impact of statins on the in vitro cytotoxicity of BCG on human urothelial carcinoma (UC) cell lines. METHODS: The human UC cell lines 253J and T24 were exposed to BCG and analyzed for upregulation of enzymes in the cholesterol biosynthe- sis pathway using Affymetrix gene expression profiling. The effect of atorva- statin on NFkB signaling and downstream gene expression, alone and in combination with BCG, was compared to non-treated and BCG exposed controls. The two cell lines were utilized to determine the effect of a physiologic concentration of atorvastatin on BCG associated cytotoxicity. RESULTS: Analysis of gene expression using functional annotation clustering demonstrated highly significant enrichment of steroid biosynthetic genes in 253J cells following BCG treatment (p = 1.4E-14). While noted, up-regulation of these genes was not significant in T24 cells (p = 0.3). Compared to both BCG exposed and untreated controls, the most pro- nounced effect of atorvastatin treatment was decreased expression of genes involved in cell cycle regulation (p  1.0e-40). This effect was independent of whether atorvastatin was used along or in combination with BCG. Atorvastatin significantly increased BCG induced signaling through the NFkB pathway (p0.001). Parodoxically, atorvastatin significantly decreased the down- stream expression of a panel of NFkB dependent, BCG induced genes as measured by quantitative rtPCR (IL-6, IL-8, CCL20, CXC1, P0.05). Expo- sure to BCG resulted in decreased proliferation of UC cells in both T24 and 253J cells lines. The addition of atorvastatin to BCG treated cells resulted in significantly decreased BCG cytotoxicity as compared to the BCG alone groups (ANOVA p = 0.001 and p = 0.003 for 253J and T24 respectively). CONCLUSIONS: Physiologic concentrations of atorvastatin exert a direct effect on the biology of urothelial carcinoma cells. In the context of BCG exposure, atorvastatin alters BCG induced signaling, inflammatory gene ex- pression, and cytotoxcity. Given a clear in-vitro effect clinicians should con- sider short term cessation of statins in patients failing to respond to an induction course of BCG. Source of Funding: Department of Veterans Affairs Vol. 183, No. 4, Supplement, Sunday, May 30, 2010 THE JOURNAL OF UROLOGY e125