l experiments, the inserts were well tolerated. No anterior chamber inflammation or corneal changes were observed after fluorescein administration in the eyes of the rats. After one week of application, insert matrix was completely mixed to the ocular mucosa. The device was able to lower IOP and completely restore the baseline values of IOP. Importantly, control glaucomatous eyes which received placebo inserts did not show any significant change in the elevated IOP. These in vivo data demonstrated that D+I reduced the IOP for up to 30 days and they were well tolerated by the animals. Of note, each chitosan insert was loaded with low amount of DIZE 13 / 18 Ocular Inserts of DIZE to Treat Glaucoma in Rats and it was effective for all experimental period. This feature is a tremendous advantage of any controlled drug delivery system because this avoids drug loss to systemic circulation since eye drops might lead to drug wastage and potential side effects. Furthermore, eye drops are susceptible to rapid tear turnover resulting in low corneal bioavailability and rapid clearance. In this way, eye drops require frequent instillation with large drug loadings to maintain the drug concentration within the therapeutic window. Noteworthy, the ocular hypotensive effect promoted by D+I did not interfere in the MAP. This characteristic is desirable for any antiglaucomatous drug or formulation as it increases its specificity reducing potential side effects. The pathophysiological rationality to explore modulators of the RAS as antiglaucomatous agents come from previous studies. For instance, the effects of blockers of the ACE/Ang II/AT1 receptor axis on IOP have been evaluated in animals and patients with glaucoma. Recently, we have proposed activation of ACE2 as a new strategy to develop drugs to treat glaucoma since this approach increases the inactivation of Ang II and get (-)-Blebbistatin production of Ang-. Indeed, our present findings showed that increased activity of endogenous ACE2 promoted by ocular inserts containing DIZE reduced the IOP of glaucomatous rats. Interestingly, 
activation of ACE2 also induces beneficial effects on uveitis. Thus, these findings, in addition to data reporting the expression of ACE2 in both human and rodent retinas and the effectiveness of DIZE as an activator of ACE2, are the rational to propose activation of intrinsic ACE2 as a new strategy to treat ocular diseases. Although we did not investigate the molecular pathways underlying the effects of DIZE, possible mechanisms such PubMed ID:http://www.ncbi.nlm.nih.gov/pubmed/19747578 as release of nitric oxide and prostaglandins induced by Ang, suppressing inflammation and inhibiting cell PubMed ID:http://www.ncbi.nlm.nih.gov/pubmed/19748727 proliferation may be involved. NO is a physiological active molecule present in rods, bipolar cells, amacrine cells and ganglion cells in retinas. It promotes relaxation of trabecular meshwork cells and ciliary muscle and its production appears to be reduced in the context of primary open-angle glaucoma. Additionally, treatment with indomethacin, an inhibitor of prostaglandins synthesis, abolished the IOP-lowering effect caused by enalaprilat, indicating that prostaglandins may mediate, at least in part, the ocular hypotensive effect of enalaprilat. Indeed, it is well known that Ang II directly induces cell proliferation and contributes to the inflammatory process by increasing the expression of pro-inflammatory cytokines, chemokines and cell adhesion molecules via AT1 receptor. Moreover, treatment with DIZE decreased the infiltration of inflammatory cells in