A Safety and Efficacy Study of 2 Dosing Regimens of Recombinant Human Nerve Growth Factor (rhNGF) Eye Drop Solution Compared With Vehicle in Patients With Dry Eye Disease.

Schirmer-I test was performed without anesthesia to determine wetting of the strip within 5 minutes, the length of the moistened strip being measured in millimeters (mm).

"Improvement" is defined as having a change from baseline at the corresponding visit >= 10 mm/5min in Schirmer-I test without anesthesia. Data analyzed and here reported were the data after having applied the predefined strategy to handle intercurrent events (ie, data for Schirmer-I Test collected in the week after an administration of a prohibited medication were set to missing under the hypothetical strategy while a treatment policy strategy was used for any other intercurrent event). Missing data for Schirmer-I Test were imputed employing an MI based on copy-reference approach assuming MNAR.

The Schirmer test was used in ophthalmic examination to measure tear production for the diagnosis of several ocular conditions such as dry eye.

Without previously instilling anesthetic drops, the Schirmer strip was inserted into the lower conjunctival sac at the junction of the lateral and middle thirds, avoiding touching the cornea, and the length of wetting strips in millimeters was recorded after 5 minutes. After 5 minutes had elapsed, the Schirmer test strip was removed and the length of the tear absorption on the strip was measured (millimeters/5 minutes).

Cutoff values:

<5 mm - pathologic dry eye 5-10 mm - marginal dry eye >10 and <30 mm - normal secretion The longer the moistened strip, the healthier the status of the eye.

Adjusted means are reported. Data here reported are the data after having applied the predefined strategy to handle intercurrent events. Missing data for Schirmer-I Test were imputed employing an MI based on copy reference approach assuming MNAR.

Corneal fluorescein staining examination was performed at the slit-lamp using blue light. Corneal fluorescein staining was graded according to the NEI scale (5 zones, each zone scoring 0-3, with a total score ranging 0-15, with a higher number representing more staining). The five cornea zones are: central, superior, inferior, nasal and temporal.

To avoid the phenomenon of quenching, which is increased in participants with dry eye, staining was assessed after 2-5 minutes from fluorescein instillation for fTBUT evaluation.

Adjusted means are reported. Data here reported are the data after having applied the predefined strategy to handle intercurrent events. Missing data for The corneal fluorescein staining (NEI score) were imputed employing an MI based on copy reference approach assuming MNAR.

Schirmer-I test was performed without anesthesia to determine wetting of the strip within 5 minutes, the length of the moistened strip being measured in millimeters (mm). The longer the moistened strip, the better the outcome.

The ones reported are adjusted means. Missing data for Schirmer-I Test were imputed employing an MI based on copy reference approach assuming MNAR.

Schirmer-I test was performed without anesthesia to determine wetting of the strip within 5 minutes, the length of the moistened strip being measured in millimeters (mm).

"Improvement" is defined as having a change from baseline at the corresponding visit >= 10 mm/5min in Schirmer-I test without anesthesia. Missing data for Schirmer-I Score were imputed employing an MI based on copy-reference approach assuming MNAR.

Samely, "Adjusted proportions" are reported, expressed under the term "mean" (the system doesn't provide the option "adjusted proportion").

fTBUT was measured by determining the time to tear film break-up. fTBUT was measured after instilling one drop of fluorescein into the inferior conjunctival cul-de-sac of the study eye. After the participant blinked several times, the examiner waited ~30 seconds. Using a slit lamp (10X, cobalt blue), the examiner recorded the fTBUT as the time from the last blink to the first break in the tear film. The shorter the time, the worse the outcome.

The fTBUT was measured twice during the first minute after the instillation. If the 2 readings differed by more than 2 seconds, then a third was taken. The fTBUT value was the average of the 2 or 3 measurements. Missing data for fluorescein tear break-up Time were imputed employing an MI based on copy-reference approach assuming MNAR. Two separate imputation models were used for each timepoint.

Adjusted means are reported.

OPAS is a validated questionnaire. Administered to all patients, it was completed only by those that either had eye pain or had filled this form before.

It has 7 domains:

• eye pain intensity over 24 hours and past two weeks
• non-eye pain intensity
• quality of life
• aggravating factors
• associated factors
• symptomatic relief Each subscale score= sum of scores divided by number of questions answered within each subscale. % values were divided by 10 before applying the algorithm.

Ocular Pain Score = sum of the scores for eye pain intensity at 24 hours and 2 weeks (questions 4-9) divided by the number of questions answered for questions 4-9. This subscore ranges 0-10, where the higher the score, the worse the outcome.

QoL score = sum of the scores for QoL subscale (questions 13-19) divided by the number of questions answered for questions 13-19. QoL scores range 0-10 where the higher the score, the better the outcome. MI was applied.

Adjusted means are reported.

Corneal fluorescein staining examination was performed at the slit-lamp using blue light. Corneal fluorescein staining was graded according to the NEI scale (5 zones, each zone scoring 0-3, with a total score ranging 0-15, with a higher number representing more staining). The five cornea zones are: central, superior, inferior, nasal and temporal.

To avoid the phenomenon of quenching, which is increased in participants with dry eye, staining was assessed after 2-5 minutes from fluorescein instillation for fTBUT evaluation. Missing data for corneal fluorescein staining were imputed employing an MI based on copy-reference approach assuming MNAR.

Adjusted means are reported.

The Symptom Assessment in Dry Eye (SANDE) questionnaire was a short questionnaire to evaluate both dry eye intensity/severity and frequency.

This questionnaire used a 100 mm horizontal line (Visual Analogue Scale - VAS) for each of the 2 questions to assess ocular discomfort and/or dryness experienced by the patients.

In the SANDE questionnaire, frequency of symptoms ranges from "rarely" to "all of the time" and the severity of symptoms ranged from "very mild" to "very severe". Patients were asked to place a mark on the two given lines based on the extent of their symptoms.

The locations of the marks was measured in mm from left to right and recorded as frequency and severity scores, respectively.

The SANDE scale ranged from 0, being the minimal amount of dry eye symptoms (best outcome) to 100, being the maximal amount of dry eye symptoms (worst outcome). In this outcome severity score was assessed. MI was applied.

Adjusted means are reported.

SANDE was a short questionnaire to evaluate both dry eye intensity and frequency. It uses a 100 mm horizontal line (Visual Analogue Scale, VAS), for each of the 2 questions, to assess ocular discomfort and/or dryness.

Frequency of symptoms ranged from "rarely" (best outcome) to "all of the time" (worst outcome), while the severity of symptoms ranged from "very mild" (best outcome) to "very severe" (worst outcome). Patients had to place a mark on the two given lines based on the extent of their symptoms.

The locations of the marks was measured in mm from left to right and recorded as frequency and severity scores, respectively.

The SANDE lines for intensity and for severity ranged from 0, being the minimal amount of dry eye symptoms (best outcome) to 100, being the maximal amount of dry eye symptoms (worst outcome). In this outcome frequency score was assessed. MI was applied.

Adjusted means are reported.

SANDE questionnaire included 2 VAS-based questions that assessed:

(i) DED symptom frequency (from 0 to 100) (ii) DED symptom severity (from 0 to 100) compiled by the participants. The global SANDE score (from 0 to 100, with 100 representing the most frequent and severe dry eye symptoms) was calculated by multiplying the frequency score by the severity score and obtaining the square root. For SANDE global Score, the lower the score, the better the outcome.

Adjusted means are reported. Data here reported are the data after having applied the predefined strategy to handle intercurrent events (i.e. data for SANDE collected in the week after an administration of a prohibited medication are set to missing under the hypothetical strategy while a treatment policy strategy is used for any other intercurrent event). Missing data for SANDE Global Score are imputed employing Multiple Imputation based on copy-reference approach assuming MNAR.

Adjusted means are reported.

The BCDVA score is measured by ETDRS letter score. The score is given by the total number of letters read at 4-m distance. If 20 or more letters are read, then the score is given by the total number of letters read + 30. If less than 20 letters are read at 4-m distance, the score is given by the sum of the letters read at 4-m distance and the letters read at 1-m distance. The equivalent logMAR score is given by:

logMAR = 1.7 - (0.02) x (ETDRS letter score). The higher the score the better the outcome. MI was applied. Adjusted means are reported.

TEAEs were defined as adverse events that were reported or worsened on or after the first dose of study treatment. Safety results are provided for overall (ocular and non-ocular) and separately for ocular and non-ocular adverse events.

TEAEs were defined as adverse events that were reported or worsened on or after the first dose of study treatment. Most frequently reported ocular adverse events (reported in > 1 participant in the total group) are reported here: at participant's level, for the "on treatment" and for the "follow-up" periods, while most frequently reported non-ocular adverse events (reported in > 1 participant in the total group) are reported at patient level for the overall period.

Corneal Endothelial Cell Density (ECD) is a measure of the number of cells per square millimeter in the innermost layer of the cornea, which is crucial for maintaining corneal transparency and hydration. ECD is typically highest at birth (around 3,000 cells/mm2) and decreases with age, reaching approximately 2,500 cells/mm2) in adulthood. A critical minimum of 400-500 cells/mm2 is required to maintain proper function, and if the density falls below this level, it can lead to corneal edema and reduced vision. Results (absolute values and change from baseline) were summarized using descriptive statistics at each scheduled visit for both eyes. Herenuder only mean change from baseline to week 8 is reported.

A summary of dilated fundoscopy (DFE) results at week 8 for the following parameters: maculopathy, posterior vitreous detachment, optic nerve abnormal appearance, retinal or virtual hemorrhage, vitritis, in the SAF is presented for the study eye and for the fellow eye.

Change from baseline in cup-to-disc ratio in the SAF population is presented for the study eye and for the fellow eye. An increased cup-to-disc ratio means the "cup" (the central depression) of the optic nerve is larger relative to the "disc" (the entire nerve head), which can indicate a loss of nerve fibers and is often a sign of glaucoma. An increase in this ratio may indicate a decrease in the quantity of healthy neuroretinal cells. For the change from baseline, only patients with a value at both baseline visit and the postbaseline visit are included.

Bulbar conjunctival redness was assessed at the slit-lamp using white light prior to vital dye instillation and graded according to the Validate Bulbar Redness (VBR 10) scale. The scale starts at grade 10 and has 10-point steps between reference images (score range 10-100). The VBR scale ranges from 10 to 100 with a higher score meaning more severe redness.

The ocular areas where the redness is assessed for the study eye and the fellow eye are:

• Nasal conjunctiva
• Temporal conjunctiva Please note that the results (absolute values and change from baseline) were summarized by conjunctiva, visit and eye using descriptive statistics.

The number and percentage of participants who discontinued study treatment due to tolerability were summarized as recorded in the eCRF EOT page.