Thursday, October 7, 2010

Abstract: BOL-303242-X

In Vivo Ocular Efficacy Profile of BOL-303242-X, a Novel Selective Glucocorticoid Receptor Agonist, In Rabbit Models of Ocular Disease.
Invest Ophthalmol Vis Sci. 2010 Sep 29. [Epub ahead of print]
Shafiee A, Bucolo C, Budzynski E, Ward KW, Lopez FJ.
Preclinical Pharmacology, Bausch & Lomb, Rochester, United States.

Purpose: To compare BOL-303242-X (formerly ZK-245186) efficacy, a novel Selective Glucocorticoid Receptor Agonist (SEGRA), to dexamethasone (DEX) in rabbit models of ocular disease. The effects of topical BOL-303242-X and DEX on intraocular pressure (IOP) and body weight changes were also evaluated.

Methods: Dry eye was induced by atropine sulfate administration, followed by treatment with saline, BOL-303242-X (0.1-1.0%), DEX (0.1%), Restasis® 0.05%, or Refresh Endura for 7-8 days. For paracentesis studies, vehicle, BOL-303242-X (0.1%, 0.5%, and 1.0%), or DEX (0.1%) were repeatedly administered topically 3 hours before paracentesis and continued for 90 min afterwards. For IOP and body weight measurements, right eyes of rabbits were topically treated with vehicle, BOL-303242-X (1.0% or 0.1%) or DEX (0.1%) four times per day for 6 weeks.

Results: In the dry eye model, BOL-303242-X and DEX were fully efficacious, maintaining tear volume and tear breakup time (TBUT) at baseline levels. Although Restasis improved tear volume versus vehicle, no changes were observed in TBUT. In the paracentesis study, BOL-303242-X and DEX improved ocular inflammation. BOL-303242-X reduced protein and PGE2 levels. Finally, BOL-303242-X showed no effects on integrated IOP or body weight, while DEX significantly increased integrated IOP and prevented the increase of body weight observed in the vehicle-treated animals.

Conclusions: BOL-303242-X shows full anti-inflammatory efficacy (similar to DEX) in experimental models of dry eye and post-operative inflammation, while demonstrating reduced effects in IOP and body weight. These data indicate that BOL-303242-X, a SEGRA, shows efficacy similar to traditional steroids, while exhibiting an improved side effect profile in IOP/muscle wasting.

Abstract: Tear collection methods for analysis

Two dimensional electrophoretic analysis of human tears: Collection method in dry eye syndrome.
Electrophoresis. 2010 Sep 29. [Epub ahead of print]
Saijyothi AV, Angayarkanni N, Syama C, Utpal T, Shweta A, Bhaskar S, Geetha IK, Vinay PS, Thennarasu M, Sivakumar RM, Prema P.
Biochemistry and Cell Biology Department, Vision Research Foundation, Sankara Nethralaya, Chennai, India.

Tear proteomics, by 2-DE, can give a fingerprint of the protein profile, which is well suited in clinical proteomics for biomarker identification and in diagnostics. The mode of tear collection can influence the representation of the proteins in the tear and therefore it is important to use the appropriate method. In this study, capillary and Schirmer mode of tear collection was done in the healthy controls and the Schirmer method was validated in dry eye syndrome conditions. 2-D PAGE of normal and dry eye tear was performed using pH 3-10 linear IPG strips followed by 13% SDS-PAGE. The spot intensity was analyzed by the PD quest software. The two methods were compared using Bland-Altman statistical tool. The 2-D map of capillary and Schirmer tear showed 147±8 spots and 145±7 spots respectively. Both the collection methods were in agreement with each other and were comparable. Dry eye tear protein showed differential expression of proteins as observed in 25-35 kDa region. One of the significantly reduced protein was identified as proline-rich 4 protein. Schirmer method of tear collection is reliable in patients with dry eye, which can display the differential protein expression and help in biomarker identification.

Abstract: Resolvin E1

Resolvin E1 Improves Tear Production and Decreases Inflammation in a Dry Eye Mouse Model.
J Ocul Pharmacol Ther. 2010 Sep 28. [Epub ahead of print]
Li N, He J, Schwartz CE, Gjorstrup P, Bazan HE.
1 Neuroscience Center of Excellence and the Department of Ophthalmology, Louisiana State University Health Sciences Center , New Orleans, Louisiana.

Purpose: Dry eye (DE) is a common ocular surface disease, particularly among women and the elderly, with chronic symptoms of eye irritation and, in severe cases, blurred vision. Several studies have shown that there is an inflammatory component in DE, although the pathogenesis is not thoroughly understood. Resolvin E1 (RvE1; RX-10001) is an endogenous mediator derived from the omega-3 polyunsaturated fatty acid eicosapentaenoic acid and is involved in inflammation resolution and tissue protection. Here we investigated the role of RvE1 in a DE mouse model.

Methods: Thirteen- to 14-week-old female BALB/C mice were exposed to desiccating conditions. One week after DE exposure, animals were treated topically with drug or vehicle 4 times per day for an additional week. Controls were nontreated animals placed in a normal environment. Schirmer's test was performed before treatment initiation and at days 2 and 4 after treatment. Density of corneal epithelial cells was analyzed in vivo using the Rostock Cornea Module of the Heidelberg Retina Tomograph (HRT-II). Corneas were processed using Western blot analysis and immunofluorescence examination.

Results: Schirmer's test showed a significant decrease in tear production in DE compared with controls. There was no change at 2 and 4 days after treatment with the vehicle, but a significant increase was observed at 2 and 4 days in the RvE1-treated group. The density of the superficial epithelial cells showed a significant decrease after DE compared with controls, which increased after 7 days of RvE1 treatment. Western blot analysis showed that α-smooth muscle actin and cyclooxygenase-2 (COX-2) expression were strongly upregulated after DE and decreased after 7 days of RvE1 treatment. Immunofluorescence confirmed strong positive staining of α-smooth muscle actin and COX-2 in stroma and/or in epithelia after DE, which decreased with RvE1 treatment. The percentage of infiltrating CD4+ T cells and CD11b+ cells decreased after RvE1 treatment when compared with DE.

Conclusion: RvE1 promotes tear production, corneal epithelial integrity, and a decrease in inflammatory inducible COX-2. In the stroma, RvE1 inhibits keratocyte transformation to myofibroblasts and lowers the number of monocytes/macrophages in this DE mouse model. These results suggest that RvE1 and similar resolvin analogs have therapeutic potential in the treatment of DE.