Aug. 6, 2009  31  US 2009/0197849 Al  Each formulation was applied in a 25 µl dosage and a 50 µl  dosage for a total of 6 test groups. The receiver chamber was  sealed to the atmosphere using parafilm wrap so that it was  spill-proof and airtight. Three diffusion cells were used for  each test group for a total of 18 cells.  [0350]  The cells were filled with DI water for a receiver  solution. The DI water had been degased to remove air  bubbles. The receiver solution was completely withdrawn  and replaced with fresh DI water at each time point. Samples  of the receiver solution were taken and analyzed by HPLC  (high pressure liquid chromatography) to determine the leu› prolide concentration. The cumulative amount of leuprolide  across human cadaver skin was calculated from a 25 µl and a  50 µl solution containing NaOH, using the measured leupro› lide concentrations in the receiver solutions for each time  point (5 and 24 hours).  solution was washed away from the skin for 3 cells ( cells #4  to 6) that were treated with 2% NaOH solution and 3 cells  (cells #13 to 15) that were treated with 4% NaOH solution.  After 24 hours, the Na OH solution was washed away from the  skinfor3 cells (cells #7 to 9) that were treated with2% NaOH  solution and 3 cells ( cells #16 to 18) that were treated with4%  Na OH solution. To wash away the Na OH solution, the receiv› ing fluid was removed and replaced with fresh DI water. This  was done twice. DI water was added to the donor chamber to  dilute the NaOH solution and then the donor solution was  removed. This was repeated several times.  [0355]  After the NaOH solution was washed away from the  skin, the solution in the donor chamber was completely  removed and replaced by 50 µl of an oxytocin solution. The  formulation of the oxytocin solution is listed in Table 45.  Once the oxytocin solution was applied, the donor chamber  was covered with parafilm.  [0351]  The cumulative amount ofleuprolide across human  cadaver skin for the 25 µl dosage at 24 hours increased from  0.52 µg/cm 2  to 4.43 µg/cm 2  when the calculated sodium  hydroxide concentration in the dried patch was increased  from 0% to 6.7%. The cumulative amount of leuprolide  across human cadaver skin for the 50 µl dosage at 24 hours  increased from 0.32 µg/cm 2  to 10.8 µg/cm $^{2 }$when the calcu› lated sodium hydroxide concentration in the leuprolide solu› tion was increased from 0% to 6.7%. The cumulative amount  ofleuprolide across human cadaver skin at 24 hours from the  50 µl  dosage group containing 3.6% NaOH (Leu-2) was 8.58  µg/cm 2 ,  which was about 27 times higher than that from the  formulation without NaOH (0.32 µg/cm 2 ,  Leu-1).  [0352]  The formulation ofLeu-2 provided up to 6-fold (25  µl solution) and to 27-fold (50 µl solution) more leuprolide  flux than in the absence ofNaOH (Leu-1 ). The formulation of  Leu-3 provided up to 9-fold (25 µl  solution) and up to 34-fold  (50 µl solution) more flux than in the absence ofNaOH.  The cells were filled with DI water as a receiver solution. The  DI water had been degased to remove air bubbles. The  receiver solution was completely withdrawn and replaced  with fresh DI water at each time point. The samples taken  were analyzed by HPLC for the concentration of oxytocin in  the receiver solution. The cumulative amount of oxytocin  across human cadaver skin was calculated using the measured  oxytocin concentrations in the receiver solutions for each  time point, which were listed in Table 46. The skin was  pretreated with 4% Na OH for the specified pretreatment time  period.  [0353]  The in-vitro permeation of oxytocin through human  cadaver skin was performed using Franz-type diffusion cells  with a diffusion area of 1 cm 2 .  The volume of receiver solu› tion was 8 ml. Human cadaver skin was cut to a proper size  and placed on a flat surface with the stratum corneum side  facing up. The skin was clamped between the donor and  receiver chambers of the diffusion cell. Eighteen diffusion  cells were used in this study. A 2% NaOH aqueous solution  (50 µl) was introduced to the donor chambers of nine cells  (cells #1 to 9) and a 4% NaOH aqueous solution (50 µl)  was  introduced to the donor chambers of the other nine cells ( cells  #10 to 18). Once the NaOH solution is applied, the donor  chamber was covered with parafilm.  [0354]  After 5 hours, the NaOH solution was washed away  from the skin for 3 cells (cells #1 to 3) that were treated with  2% NaOH solution and 3 cells (cells #10 to 12) that were  treated with 4% NaOH solution. After 10 hours, the NaOH  [0356]  The in-vitro permeation of oxytocin through human  cadaver skin was performed as described in Example 14,  except that a 0.25% NaOH aqueous solution (50 µl) was  introduced to the donor chambers of nine cells ( cells #1 to 9)  and a 1.0% NaOH aqueous solution (50 µl)  was introduced to  the donor chambers of the other nine cells (cells #10 to 18).  [0357]  After 5 hours, the NaOH solution was washed away  from the skin for 3 cells (cells #1 to 3) that were treated with  0.5% NaOH solution and 3 cells (cells #10 to 12) that were  treated with 1.0% NaOH solution. After 11 hours, the NaOH  solution was washed away from the skin for 3 cells ( cells #4  to 6) that were treated with 0.25% NaOH solution and 3 cells  ( cells #13 to 15) that were treated with 1.0% NaOH solution.  After 24 hours, the Na OH solution was washed away from the  TABLE46  TABLE45  TABLE 44  Example 15  Example 14  Cumulative Amount ofOxytocin (l!g/cm 2 )  Time  5 hr Pretreatment  15 hr Pretreatment  24 hr Pretreatment  5 hours  118.95  202.28  193.82  15 hours  200.66  222.45  232.72  24 hours  225.52  231.58  236.80  Formulation for the Oxytocin Solution  Ingredient  g  Oxytocin  0.005  DI water  0.6  PG  0.6  Cumulative Amount ofLeu12rolide (l!g/cm 2 )  25 l!l solution  50 l!l solution  Time  Leu-1  Leu-2  Leu-3  Leu-1  Leu-2  Leu-3  5 hours  0.38  0.52  0.58  0.32  0.62  0.3  24 hours  0.52  3.21  4.43  0.32  8.58  10.8 