Effect of electroporation and pH on the iontophoretic transdermal delivery of human insulin.
The synergistic effect of electroporation (EP) and iontophoresis (IP) on the in vivo percutaneous absorption of human insulin was evaluated in rats. Passive diffusion and IP alone (0.4 mA/cm(2)) resulted in almost no skin permeation of insulin at pH 7, whereas EP treatment (150 or 300 V, 10 ms, and 10 pulses) resulted in a high plasma level of insulin and the combined use of EP and IP led to a further increase of the plasma level of insulin compared with that measured after EP alone. Interestingly, a much higher plasma level was observed when the pH of the insulin solution at 7 was increased to 10. One of the reasons was the different aggregation properties of insulin at pH 7 and pH 10. The nonassociation ratio of insulin was significantly higher at pH 10 than at pH 7. Insulin monomers and dimers were observed in addition to the normal form of insulin, hexamer, albeit in low percentages, at pH 10, whereas most of the insulin was in the hexamer form at pH 7. To confirm the influence of the aggregation properties of insulin, the commercially available human insulin analogue insulin lispro was then evaluated. Its skin permeation was found to be extremely high compared to that of conventional human insulin without increasing the solution pH. Marked decreases in blood glucose levels reflecting the increases in the plasma concentration of insulin were also observed after EP/IP treatment. The present study suggests that percutaneous absorption of insulin is synergistically enhanced by a combined use of EP and IP and that altering the aggregation properties of insulin is important to enhance the percutaneous absorption of insulin by IP and/or EP.