Safety assessments included AEs, ocular examination, spirometry, safety laboratory tests (hematology, serum chemistry, and urinalysis), 12-lead and telemetry ECGs, vital signs including respiratory rate, and physical examinations. We recorded AEs from day 1 post dose through the follow-up visit on day 7. Ocular safety was investigated by slit-lamp examination, including assessment of injection, cells and flare, slit-lamp examination with indirect ophthalmoscopy for examination of the fundus, fundus photomicrography, BCVA by Early Treatment Diabetic Retinopathy Study chart, and pupillometry. Fundus imaging was obtained before and after spirometry, to ensure that no retinal vascular changes occurred as a result of either the drug or spirometry. Subjects underwent dilated fundus examinations with infrared fundus photographic imaging (Heidelberg Spectralis; Heidelberg Engineering, Heidelberg, Germany) on day 2 (before spirometry); day 1 predose (after spirometry); and at 5, 10, and 30 minutes, and 2, 5, and 73 hours after administration of study drug. Image analysis was by central reading which was conducted by the same independent, masked reader. Retinal vasculature changes were evaluated by dilated fundus photomicroscopy and indirect ophthalmoscopy examination. Symptoms related to retinal vasoconstriction were assessed (photopsias, zig zag image of light patterns, fortification pattern of lights, and transient obscuration of vision). Indirect ophthalmoscopy was used for evaluation of the retina.
Ocular tolerability was evaluated by subject ratings of ocular discomfort (photophobia, itching, tearing, dryness, and discharge, rated on a scale of 0–4) and investigator rating of conjunctival hyperemia using an ocular redness scale (Ora Calibra #6.0b; used under license from Ora, Inc., Andover, MA, USA). Patients were rated on a scale of 0 to 3 (0–0.5: absent; 1–1.5: mild; 2–2.5: moderate; and 3: severe). Scores were compared to baseline and analysis of hyperemia was based on scores taken from the worst eye.