OPHTHALMIC PREPARATIONS

Abstract: An ophthalmic preparation contains human epidermal growth factor at a concentration of 0.01 to 50 µg/ml (or g), preferably 0.5 to 30 µg/ml (or g) and a diluent such as a buffer solution (pH 5.0 - 8.0) or an ointment base. The preparation is effective to cure keratitis, corneal erosion, corneal infiltration or corneal ulcer.

Description: Ophthalmic preparations This invention relates to an ophthalmic preparation containing human epidermal growth factor (hereinafter referred to as h-EGF) as an active component. ore particularly, the invention relates to an ophthalmic preparation which is of value in the treatment of keratitis, corneal erosion, corneal infiltration and corneal ulcer. h-EGF, which is derived from human urine, is a straight-chain polypeptide having 3 disulfide bridges and a molecular weight of about 6,000. This substance has epitheliogenic activity. Regarding the action mechanism of h-EGF it has been suggested that the substance first binds itself to the EGF receptor on the cell membrane, then finds its way into the cell to induce DNA synthesis and, hence, proliferation of the cell [Journal of the Japan Medical Association 85, (7) 833, 1981]. As to the tissue distribution of h-EGF, it occurs in the saliva, blood and urine at physiologically effective levels but does not occur in any appreciable amount in the lacrimal fluid, i.e. tear. In local administration of h-EGF to patients with corneal injury, the present inventors found that the injuries were cured very soon after the treatment. On the other hand, animal experiments and a trial in healthy subjects, which will hereinafter be quoted, revealed that the intact corneal cell is not influenced by the action of h-EGF. After administration of h-EGF to mice and rats in groups of 6 individuals of either sex at the dose level of 10 mg/kg by the subcutaneous route and at the dose of 1 mg/kg by the intravenous route (the doses equivalent to about 1,000,000 and about 100,000 times the amount in the blood of human/kg) caused no change in general condition, nor death, indicating that it would not produce any toxic effects. Another experiment in which a 5 pgtml solution of h-EGF was administered to male and female rabbits in groups of 3 individuals of either sex at the dose of 3 drops per instillation at the frequency of 3 times daily for 1 month showed that it caused no abnormalities in the ocular membrane. It was therefore found that h-EGF is a polypeptide which has only a very low toxicity. This invention, which has thus been based on the above findings, is directed to an ophthalmic preparation which comprises an effective amount of human epidermal growth factor and an ophthalmologically acceptable diluent. The ophthalmic preparation of h-EGF can be made available in such dosage form as an ophthalmic lotion, an ophthalmic ointment, etc. The concentration of h-EGF in such an ophthalmic preparation is preferably in the range of 0.01 to 50 pg/ml (or g). In said ophthalr-ic lotion, water or an aqueous buffer solution may be used as a diluent. The buffer used for pH adjustment in the preparation of this invention may be any buffer substance that is ophthalmologically acceptable. Thus, for example, such a combination as sodium chloride, monopotassium phosphate, potassium chloride, calcium chloride and magnesium sulfate; sodium dihydrogen phosphate and disodiu hydrogen phosphate; or sodium chloride, potassium chloride and calcium chloride may be used. For the preparation of said ophthalmic ointment, a diluent (base) such as white vaseline, purified lanoline, liquid paraffin and the like may be employed. To prepare the ophthalmic ointment, h-EGF may for instance be directly mixed with such a base or alternatively first dissolved in water, then mixed with lanolin and finally mixed with a further amount of lanolin or another ointment base. In addition to these procedures, any other established pharmaceutical procedure for the preparation of ophthalmic ointments may be employed, if desired. Furthermore, in the ophthalmic preparation according to this invention, there may also be incorporated any of such conventional preservatives as chlorobutanol, methyl p-oxybenzoate, benzyl alcohol, etc. in addition to said pH-adjusting buffers. The pH of the ophthalmic preparation need only within the ophthalmolcgically acceptable range of pH 5.0 to 8.0 and the preparation may be prepared by subjecting the composition to sterilization by millipore filtration within the pH range. h-EGF is stable in the above pH range. The following Examples and Test Example are further illustrative of the preparation of this invention. Example 1 In sterile pure water were dissolved 0.835 w/v% of sodium chloride, 0.007 w/v % of monopotassium phosphate, 0.022 w/v % of potassium chloride, 0.017 w/v % of calcium chloride, 0.030 w.v % of magnesium sulfate. Then, h-EGF was added to portions of this solution to prepare liquids containing h-EGF in the concentrations of 0.5 pg/ml, 1.5 pg/ml and 4.5 Ug/ml, respectively. Finally, 0.15 % of chlorobutanol was added to the above liquids and the entire compositions were subjected to millipore filtration to give ophthalmic lotions. Example 2 In sterile distilled water were dissolved 0.825 w/v % of sodium chloride, 0.145 w.v % of potassium chloride and 0.015 w/v % of calcium chloride. Then, h-EGF was added to portions of the above solution in the concentration of 10 pg/ml, 15 pg/ml and 30 ug/ml, respectively followed by bacterial filtration to give ophthalmic lotions. Example 3 In a small amount of pure water was dissolved 100 g of h-EGF and the solution was thoroughlykneaded with 10 g of purified lanolin and, then, with a further 100 g of white lanolin to give an ophthalmic ointment. Test Example (1) The 10 Dg/ml, 15 pg/ml and 30 pg/ml ophthalmic lotions prepared in Example 2 were administered to male and female rabbits in groups of 3 individuals of either sex in the dose of 3 drops per instillation at the frequency of 3 times daily for 1 month to investigate the influence of the drugs on the ocular membrane. There were no abnormal findings. (2) Using 2 rabbits, uniform corneal injuries were produced in both eyes of each rabbit with an electric iron and the 1.5 ijg/mlophthalmic lotion of Example 1 was instilled into one eye and physiological saline into the fellow eye in dose of 3 to 4 drops at the frequency of 3 times daily to investigate the healing process. The results are shown in Table 1. Table l Time to cure, in days Rabbit No. H-EGF Saline 1.5 ug/ml 1 3 days 9 days 2 4 days 10 days (3) To healthy subjects, the 0.5 ug/ml, 1.5 ug/ml and 4.5 ug/ml ophthalmic lotions of Example 1 were administered in the dose of 3 to 4 at the frequency of 4 times daily for 1 week. There were no abnormal findings. Example 4 The 1.5 pg/ml ophthalmic lotion of Example 1 was administered to patients with keratitis, corneal erosion, corneal infiltration or corneal ulcer in the dose of 3 to 4 drops per instillation at the frequency of 4 times daily. The results are shown in Table 2. The time to complete cure was as short as about 1/2 of the tme period hitherto experienced, showing that the drug produces an early cure, especially in cases of corneal trauma due to contact lenses. Table 2 Disease No. of effec- Time to complete tive cases cure (in days) eratitis 18/20 2-6 days Corneal erosion 20/20 1-5 days Corneal infiltration 15/15 4-5 days Corneal ulcer 8/15 7-14 days Six cases sampled randomly from among the above cases, will be described below in further detail. Case 1. A 34-year-old male. Right eye. Corneal erosion (corneal burn) Hot water entered into the right eye and the epithelium of the entire cornea was thermally ablated. -%t visit, pain (+++), ciliary hyperemia (+++), and visual acuity 0.01. The patient was instructed to instill to 4 drops of a 1.5 pg/ml h-EGF ophthalmic lotion -nto the eye four times daily. On the following day, the visual acuity was restored. Pain and ciliary hyperemia were improved to (+) and (-), repectively. The topical application of the drug was continued for one additional day. When he visited the hospital on the next day, the cornea was quite clear and the subjective symtoms had disappeared. Case 2. A 32-year-old male. Left eye. Corneal erosion. A synthetic resin adhesive agent (cyanoacrylate) having entered into his left eye, the patient visited the hospital. Immediately, foreign bodies were eliminated. The elipthelium of the entire cornea had become detached; pain (+++), ciliary hypermia (++), and visual acuity 0.06. The instillation of a 1.5 pg/ml h-EGF ophthalmic lotion was instituted (3 to 4 drops, four times daily). On the following day, pain was slightly improved (++) but epithelial ablation and ciliary hyperemia (++) remained unchanged. On the fourth day, erosion became very slight but pain (+) and ciliary hyperemia (++) persisted. The patient was instructed to continue the topical application of the h-EGF ophthalmic lotion, and on the sixth day, the cornea became quite clear and the subjective symptoms completely disappeared. Visual acuity recovered to 1. 0. Case 3. a 82-year-old female. Left eye. Corneal ulcer. Ulcer occurred in the area of previous parenchymatous keratitis, resulting in pain (++), ciliary hyperemia (++) and severe visual disorder. Instillation of a 1.5 Vg/ml h-EGF ophthalmic lotion was instituted. The dosage was 3 to 4 drops four times daily. On the fourth day, pain and ciliary hyperemia improved to (+) and (i), respectively. On the seventh day, pain and ciliary hyperemia both improved to (-), but slight ulcer persisted in the central area. In the eleventh day, the cornea was stained only very faintly on fluorescein staining. By the fifteenth day, the corneal ulcer had completely healed and the visual acuity recovered to 30 cm/n.d. Case 4. A 61-year-old male. Right eye. Corneal infiltration. The patient visited the hospital because of onset of moderate corneal infiltration. Pain (+), ciliary hyperemia (++), and visual acuity 0.5. Topical therapy with 3 to 4 drops of a 1.5 pg/ml h-EGF ophthalmic lotion four times daily was instituted. Consequently, by the seventh day, the infiltration had subsided almost completely and the subjective symptoms disappeared, and the instillation of the drug was discontinued. However, 5 days later, infiltration relapsed and the h-EGF ophthalmic therapy was reinstitu ted according to the same dosage regimen. On the 1Oth day after reinststaticn both the infiltration and subjective symptoms disappeared. No recurrence was found thereafter. Visual acuity was 0.5. Case 5. A 67-year-old male. Both eyes. Keratitis. Keratitis occurred in the surface stratum of corneal macula, of which the patient had a past history. Pain (+), ciliary hyperemia (+), and visual acuity 0.09, right and 0.03, left. Three to four drops of the 1.5 pg/ml h-EGF ophthalmic lotion were instilled into the eye four times daily. By the fourth day, both the corneal inflammation and subjective symptoms had almost disappeared. On the sixth day, the keratitis was completely cured. The visual acuity was 0.2, right and 0.09, left. Case 6. A 38-year-old male. Right eye. Corneal ulcer. Corneal ulcer developed around the pseudopterygium. Pain (+), photophobia (++), ciliary hyperemia (++), foreign body sensation (++), and visual acuity 1.2. h-EGF ophthalmic lotion, 1.5 ug/ml, (3 to 4 drops four times daily) was topically applied. On the third day, the epithelium began to cover the ulcerative area and all the subjective symptom was foreign body sensation. On the tenth day, the cornea ceased to be stained with flucrescein and the subjective symptom disappeared. Visual acuity = 1.5.

Claims: CLAIMS: 1. An ophthalmic preparation characterised in that it comprises an effective amount of human epidermal growth factor. 2. A preparation as claimed in Claim 1, wherein the human epidermal growth factor is contained at a concentration of 0.01 to 50 pglml (or g). 3. A preparation as claimed in Claim 2 wherein said concentration is 0.5 to 30 pg/ml (ore). 4. A preparation as claimed in Claim 3, wherein the said concentration is 0.5 to 4.5 ,ug/ml (or g). 5. A preparation as claimed in any one of the preceding Claims, wherein the preparation is an ophthalmic lotion. 6. A preparation as claimed in Claim 5 wherein human epidermal growth factor is diluted with an aqueous buffer solution. 7. A preparation as claimed in any one of Claims 1 to 4, wherein the preparation is an ophthalmic ointment. 8. A preparation as claimed in Claim 7, wherein human epidermal growth factor is diluted with vaseline, lanoline or liquid paraffin. 9. A preparation as claimed in claim 7 or Claim 8, wherein the preparation contains about 10 rg/g of human epidermal growth factor. 10. Human epidermal growth factor for use in the treatment of keratitis, corneal erosion, corneal infiltration or corneal ulcer.

Inventors: ASAOKA, MAKOTO, HIRATANI, HAJIME
Applicants: J C R KABUSHIKI KAISHA ALSO KNOWN AS  J C R INCORPORATED
Application year: 1983
Main classification: A61K35
IPC codes: A61K38