Safety and Efficacy of rhNGF Eye Drops at Different Doses in Patients With Dry Eye

The SANDE is a short questionnaire to evaluate the frequency of ocular dryness and/or irritation symptoms. For the assessment, the patients mark on a 100 mm Visual Analogue Scale (VAS) line the point that they feel represents their perception of their current state. The VAS score is determined by measuring in millimetres from the left hand end of the line to the point that the patient marks. The SANDE scores (0-100) will be then evaluated for frequency per day.

Time frame: Baseline, Day 1, Day 8, Day 29 and Day 56

Population: FAS subjects: all enrolled patients, who received at least one dose of the IMP. This analysis set was used for the efficacy analysis

The cornea is divided into five sectors (central, superior, inferior, nasal and temporal), each of which is scored on a scale of 0-3, with a maximal score of 15. Both nasally and temporally, the conjunctiva is divided into a superior paralimbal area, an inferior paralimbal area and a peripheral area with a grading scale of 0-3 and with a maximal score of 9 for the nasal and temporal conjunctiva. Staining was derived as the sum of scores in the various sectors. The higher the total score the more compromised is the ocular surface.

Time frame: Baseline, Day 1, Day 8, Day 29 and Day 56

Population: FAS subjects: all enrolled patients, who received at least one dose of the IMP. This analysis set was used for the efficacy analysis

The SANDE is a short questionnaire to evaluate the severity of ocular dryness and/or irritation symptoms. For the assessment, the patients mark on the 100 mm VAS line the point that they feel represents their perception of their current state. The VAS score is determined by measuring in millimetres from the left-hand end of the line to the point that the patient marks. The SANDE scores (0-100) will be then evaluated for severity. The higher the score, the worse the outcome.

Time frame: Baseline, Day 1, Day 8, Day 29 and Day 56

Population: FAS subjects: all enrolled patients, who received at least one dose of the IMP. This analysis set was used for the efficacy analysis

The Schirmer test type I (without anaesthesia) was performed to measure aqueous tear secretion prior to the instillation of any dilating or anaesthetic eye drops. The rounded tip of a standardized paper strip is inserted into the lower fornix of the eye, and the wetted length extending out from the lower lid is recorded after 5 min of eye closure. Both eyes could be tested at the same time. Changes from baseline in values of Schirmer's test type I are summarised by eye and evaluation visit, and stratified by severity level. The longer the wetted length the healthier the status of the eye. Only study eye's results are reported hereunder.

Time frame: Baseline, Day 1, Day 8, Day 29 and Day 56

Population: FAS subjects:

The treatment-emergent adverse events were recorded throughout the whole study.

Time frame: Throughout the study up to day 56

Population: Safety set: all patients who received at least one dose of the IMP. This analysis set was used for the safety analysis

Four conjunctival impression cytology samples (temporal, nasal, inferior and superior bulbar conjunctiva) for conjunctival goblet cell counts were performed in the worse eye (study eye). Conjunctival epithelium samples are obtained following the instillation of a preservative-free anaesthetic eye drop by slightly pressing on the bulbar conjunctiva a 0.1 μm cellulose acetate filter. When the disc is removed the apical layers of conjunctival epithelium remain impressed on it. Cells on the filter are fixed and stained. The final results will be expressed as mean ± SD of 3 consecutive optic fields for each sample. A higher number of goblet cells indicates a healthier eye.

Time frame: Baseline, Day 1, Day 8, Day 29 and Day 56

Population: FAS: all enrolled patients, who received at least one dose of the IMP. This analysis set was used for the efficacy analysis.

Fluorescein staining of the cornea is a methodology to visualize corneal epithelial defects under slit lamp microscopy in patients with suspicious or known Dry Eye Disease (DED). Fluorescein dyed - impregnated paper strips were used. Before placing the strip in the lower fornix of the eye, a drop of sterile saline was added to the strip. The cornea was divided into five sectors (central, superior, inferior, nasal and temporal), each of which was scored on a scale of 0-3, with a maximal global score of 15. For a better reading under the slit lamp, no intense illumination beam was used, since it could reduce the contrast and lead to an underestimation of grading. The lower the score the lower the corneal damage.

Time frame: Baseline, Day 1, Day 8, Day 29 and Day 56

Population: FAS: all enrolled patients, who received at least one dose of the IMP. This analysis set was used for the efficacy analysis.

Corneal sensitivity was measured in cm through the Luneau-Cochet-Bonnet aesthesiometer. This contains a thin, retractable, nylon monofilament that extends up to 6 cm in length. Variable pressure can be applied to the cornea by adjusting the monofilament length. The monofilament length ranges from 6 to 0.5 cm. As the monofilament length is decreased the pressure increases from 11 mm/g to 200 mm/g. The filament is retracted incrementally in 0.5 cm until the patient gives a positive reaction indicating that the contact of the monofilament on the cornea has been sensed. The shorter filament lengths indicate decreased corneal sensation. The length of the filament (in cm) at which the patient sensed the contact with the cornea is recorded. Only study eye's results are reported hereunder.

Time frame: Baseline, Day 1, Day 8, Day 29 and Day 56

Population: FAS: all enrolled patients, who received at least one dose of the IMP. This analysis set was used for the efficacy analysis.

IOP was performed using either Goldmann applanation tonometry or a handheld applanation tonometer (e.g. Tonopen) after the instillation of a topical anaesthetic. IOP was measured in both eyes after completion of all other slit lamp examinations to avoid potential interference with the other evaluations. Only study eye's results are reported hereunder.

Time frame: Baseline, Day 1, Day 8, Day 29 and Day 56

Population: FAS: all enrolled patients, who received at least one dose of the IMP. This analysis set was used for the efficacy analysis.

The OSDI is based on a 12-item questionnaire designed to provide a rapid assessment of the symptoms of ocular irritation consistent with dry eye disease and their impact on vision-related functioning. The 12 items of the OSDI questionnaire are graded on a 0-4 scale as follows: Grade 0 = none Grade 1 = some Grade 2 = half Grade 3 = most Grade 4 = all The total OSDI score was then calculated on the basis of the following formula: OSDI=\[(sum of scores for all questions answered) × 100\]/\[(total number of questions answered) × 4\]. Thus, the OSDI total score scales from 0 to 100, with higher scores representing greater disability.

Time frame: Baseline, Day 1, Day 8, Day 29 and Day 56

Population: FAS: all enrolled patients, who received at least one dose of the IMP. This analysis set was used for the efficacy analysis.

A ocular tolerability score was determined using a 100 mm VAS on which 0 meant No symptoms and 100 meant the Worst possible discomfort. The patients subjectively evaluated their ocular symptoms (foreign body sensation, burning or stinging, itching, pain, sticky feeling, blurred vision and photophobia) using the VAS giving the value they were feeling from none to an extreme value. The ocular symptoms were evaluated by the patients through the scale. Only the study eye's results are reported hereunder.

Time frame: Baseline, Day 1, Day 8, Day 29 and Day 56

Population: FAS: all enrolled patients, who received at least one dose of the IMP. This analysis set was used for the efficacy analysis

SLE grading of the eyelids, lashes, conjunctiva, cornea, lens, iris and anterior chamber was done according to the following scales: Eyelid - Meibomian glands (evaluation of the central ten Meibomian gland openings in the mid-portion of the upper eyelid): 0, 1, 2, 3 = None, Mild, Moderate, Severe gland plugging Eyelid - Erythema 0, 1, 2, 3,4 = None, Mild, Moderate, Severe, Very severe redness of lid margin and/or skin Eyelid - Oedema 0, 1, 2, 3,4 = None, Mild, Moderate, Severe, Very severe oedema Lashes 0 = Normal 1 = Abnormal Conjunctiva - Erythema 0, 1, 2, 3, 4 = None, Mild, Moderate, Severe erythema Conjunctiva - Oedema 0, 1, 2, 3, 4 = None, Mild, Moderate, Severe, Very severe swelling Lens 0, 1, 2, 3 = No, Mild, Moderate, Severe opacification N/A = Patient with artificial lens Iris 0 = Normal 1 = Abnormal. Anterior Chamber Inflammation 0, 1, 2, 3, = No, Mild, Moderate, Severe, Very severe Tyndall effect Only the study eye's results are reported hereunder.

Time frame: Baseline, Day 1, Day 8, Da 29 and Day 56

Population: FAS: all enrolled patients, who received at least one dose of the IMP. This analysis set was used for the efficacy analysis

TFBUT was measured by determining the time to tear break-up. The TFBUT was performed after instillation of 5 μl of 2% preservative-free sodium fluorescein solution into the inferior conjunctival cul-de-sac of each eye. With the aid of a slit lamp at 10X magnification using cobalt blue illumination, the examiner will monitor the integrity of the tear film, noting the time it takes to form lacunae (clear spaces in the tear film) from the time that the eye is opened after the last blink. The longer the time the better the integrity of the tear film. Only Study eye's results are reported hereunder.

Time frame: Baseline, Day 1, Day 8, Day 29 and Day 56

Population: FAS: all enrolled patients, who received at least one dose of the IMP. This analysis set was used for the efficacy analysis

Values of tear film osmolarity and their changes from baseline (screening visit) are summarised by eye (study eye and non study eye) and evaluation visit and stratified by severity level. Only study eye's results are reported hereunder.

Time frame: Changes from baseline up to day 56±4

Population: FAS: all enrolled patients, who received at least one dose of the IMP. This analysis set was used for the efficacy analysis.

The Schirmer test type II (with anaesthesia) was performed to measure aqueous tear secretion following the instillation of a preservative-free anaesthetic eye drop (Oxybuprocaine Chlorhydrate 0.4%). The rounded tip of a standardized paper strip is inserted into the lower fornix of the eye, and the wetted length extending out from the lower lid is recorded after 5 min of eye closure. Both eyes could be tested at the same time. Changes from baseline in values of Schirmer's test type I are summarised by eye and evaluation visit and stratified by severity level. The longer the wetted length the healthier the status of the eye. Only study eye's results are reported hereunder.

Time frame: Baseline, Day 1, Day 8, Day 29 and Day 56

Population: FAS: all enrolled patients, who received at least one dose of the IMP. This analysis set was used for the efficacy analysis.

Values of Best-Corrected Distance Visual Acuity (BCDVA) scores were measured using the Early Treatment Diabetic Retinopathy Study (ETDRS) score. The ETDRS charts use letters, or a geometric progression in letter size from line to line, under standardized lighting conditions. The patient starts at the top of the chart, or on the last row where he or she can read all of the letters, and reads down the chart until he or she reaches a row where a minimum of three letters on a line cannot be read. The patient is scored by how many letters could be correctly identified. Therefore, the higher the number of letters the higher the visual acuity. Changes in the ETDRS score from baseline (screening visit) are summarised by eye (study eye and non study eye) and evaluation visit, and stratified by severity level. Only study eye's results are reported hereunder.

Time frame: Baseline, Day 1, Day 8, Day 29 and Day 56

Population: FAS: all enrolled patients, who received at least one dose of the IMP. This analysis set was used for the efficacy analysis.

During the treatment with rhNGF at both doses (from day 1 to day 29), and in the follow-up period (from day 29 to day 56) the mean frequency of daily use of artificial tears was measured.

Time frame: Day 1-Day 8, Day 9-Day 29, Day 30-Day 56 intervals

Population: FAS: all enrolled patients, who received at least one dose of the IMP. This analysis set was used for the efficacy analysis.

This test allows seeing inside the fundus of the eye and other structures using an ophthalmoscope. The fundus examination included assessments of vitreous, macula, retina and optic nerve head for both eyes. Only the results concerning the study eye are reported hereunder. These structures will be assessed according to the criteria outlined below. Vitreous The examiner will judge the appearance of the vitreous in the visual axis. Normal: Absence of any opacity Abnormal: Presence of opacity Macula, (Peripheral) Retina and Optic Nerve Head The examiner will provide a separate assessment of the macular, choroid and peripheral retina Normal: Absence of any structural or vascular change, inflammation, oedema or haemorrhage. Abnormal: Evidence of any ongoing or previous structural/vascular change, inflammation, oedema or haemorrhage.

Time frame: Baseline, Day 1, Day 8, Day 29 and Day 56

Population: FAS: all enrolled patients, who received at least one dose of the IMP. This analysis set was used for the efficacy analysis.