Korean J Ophthalmol.  2016 Aug;30(4):302-308. 10.3341/kjo.2016.30.4.302.

The Effect of TNF-α Blocker HL036337 and Its Best Concentration to Inhibit Dry Eye Inflammation

• Choi W
• Noh H
• Yeo A
• Jang H
• Ahn HK
• Song YJ
• Lee HK

• KMID: 2373991
• DOI: http://doi.org/10.3341/kjo.2016.30.4.302

Abstract

PURPOSE
Dry eye syndrome is commonly thought of as an inflammatory disease, and we have previously presented data showing the effectiveness of topical TNF-α blocker agents for the treatment of this condition. The purpose of this study was to investigate the effectiveness of the TNF-α blocking agent HL036337 compared to cyclosporine A for the treatment of dry eye induced inflammation in order to establish whether HL036337 represents a more effective method for suppressing inflammation. The efficacy of HL036337 and cyclosporine A was determined using an experimental murine dry eye model.
METHODS
The TNF-α blocker HL036337 is a modified form of TNF receptor I. Using dry eye induced C57BL/6 mice (n = 45), corneal erosion was measured at day 4 and 7 after topical treatment with cyclosporine A or HL036337. To determine the effective treatment dose, 0.25, 0.5, 1, 2.5, and 5 mg/mL of HL036337 were topically administered twice per day to dry eye induced murine corneas for 1 week.
RESULTS
The optimal concentration of the TNF-α blocker HL036337 for treatment of dry eye induced corneal erosion was determined to be 1 mg/mL. Dry eye induced corneal erosion was improved after 1 week with topically applied cyclosporine A and HL036337 at 1 mg/mL.
CONCLUSIONS
HL036337 administered topically at 1 mg/mL effectively improved corneal erosion induced by dry eye. This finding may also suggest that inhibition of TNF-α can improve dry eye syndrome.

Keyword

Corneal erosion; Dry eye; HL036337; Inflammation; Tumor necrosis factor-alpha

MeSH Terms

Animals
Cornea/diagnostic imaging
Disease Models, Animal
Dose-Response Relationship, Drug
Dry Eye Syndromes/diagnosis/*drug therapy
Female
Mice
Mice, Inbred C57BL
Microscopy, Acoustic
Ophthalmic Solutions/administration & dosage
Tumor Necrosis Factor-alpha/*antagonists & inhibitors
Ophthalmic Solutions
Tumor Necrosis Factor-alpha

Figure

• 

  Fig. 1 Corneal erosion was measured followed by fluorescein cornea surface staining. Each group included six to eight corneas randomly selected from both eyes dry eye (DE) induction was performed for 10 days, after which corneal erosion in DE induced mice was measured and photographed. After 7 days of treatment with topical HL036337, corneal fluorescein staining was examined with a slit lamp biomicroscope and photos were taken. (A) Treatment with HL036337 0.25 mg/mL, (B) HL036337 0.5 mg/mL, (C) HL036337 1 mg/mL, (D) HL036337 2.5 mg/mL, and (E) HL036337 5 mg/mL. Representative photographs from each group are shown.

• 

  Fig. 2 After induction of dry eye (DE) for 10 days, topical eye drops were given to the mice groups and corneal fluorescein staining was examined with a slit lamp biomicroscope at day 0 , 4, and 7. The mice were then divided into groups as follows: DE induced, DE + HL036337 0.25 mg/mL (H 0.25), DE + HL036337 0.5 mg/mL (H 0.5), DE + HL036337 1 mg/mL (H 1.0), DE + HL036337 2.5 mg/mL (H 2.5), DE + HL036337 5 mg/mL (H 5.0), DE + Restasis, and DE + vehicle (D + V). Punctate staining was evaluated in a masked fashion using the Oxford Scheme grading system, where a grade of 0 to 4 was given for each cornea. Each experiment was repeated three times. Student's t-test, **p < 0.05. (A) Corneal erosion grading before topical eye drops treatment. (B) Corneal erosion grading after 4 days of topical eye drops treatment. (C) Corneal erosion grading after 7 days of topical eye drops treatment.

• 

  Fig. 3 Determination of clinical data. Corneal erosion treatment was evaluated for a week with topical application and compared to that of Restasis. Punctate staining was evaluated in a masked fashion using the Oxford Scheme grading system, where a grade of 0 to 4 was given for each cornea. Each experiment was repeated three times. DE = dry eye. p < 0.05.

• 

  Fig. 4 After 10 days of dry eye (DE) induction, topical eye drops were given to the mice groups and inflammatory cytokines in corneas and lymph nodes (LNs) were determined by quantitative reverse transcription polymerase chain reaction at day 7. Groups were divided as follows: normal (N), DE + vehicle (D + V), DE + HL036337 0.25 mg/mL (H 0.25), DE + HL036337 1 mg/mL (H 1.0), and DE + HL036337 2.5 mg/mL (H 2.5). Six corneas and six to eight cervical LNs from five mice were included in each group. Each experiment was repeated three times. *p < 0.05.

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