SOLUTION CONTAINING LYSOZYME HYDROCHLORIDE AND DIPOTASSIUM GLYCYRRHIZINATE

Abstract: An aqueous solution comprises lysozyme hydrochloride, dipotassium glycyrrhizinate, at least one selected from the group consisting of sodium chloride, potassium chloride, calcium chloride, magnesium chloride, sodium phosphate, potassium phosphate, sodium citrate, potassium citrate, sodium carboante and potassium carbonate and water, has a pH of 5 to 9 and is useful for eye drops.

Description: The present invention relates to a solution containing lysozyme hydrochloride and dipotassium glycyrrhizinate. In particular, the present invention relates to a solution useful in the fields of medicines and cosmetics. [Prior Art ] Lysozyme hydrochloride, being an enzyme capable of decomposing mucopolysaccharides contained in various mucuses secreted from tissues, decomposes cell walls of bacteria to show its bacteriolytic effects in an infected patient. When the patient is suffering from inflammation, a membranous mucus produced by the inflammation is decomposed to facilitate the excretion thereof and to exhibit an antiphlogistic effect. Thus, lysozyme hydrochloride is frequently and effectively used when a mucous membrane of eyes, nose, throat or ears is inflamed by an infection. Lysozyme hydrochloride is contained in not only preparations to be administered perorally but also those to be directly applied to an affected part in the form of aqueous solution. A preferred example of the formulations is eye drops which have already been put on the market as a remedy for chronic conjunctivitis. Dipotassium glycyrrhizinate has an antiallergic effect and it is useful as a so-called non-steroidal antiinflammatory agent. Its antiinflammatory effect is remarkable, particularly on an allergic inflammation of a mucous membrane. Therefore, medicines containing dipotassium glycyrrhizinate were already put on the market as eye drops to be directly applied in the treatment of, for example, allergic conjunctivitis and allergic blepharitis. As described above, lysozyme hydrochloride and dipotassium glycyrrhizinate have been used independently from each other as remedies for inflammations of mucous membranes. It is expected, however, that the therapeutic effects are further improved when a combination of them can be used. Namely, it is desired to provide a solution of a mixture of lysozyme hydrochloride and dipotassium glycyrrhizinate to be directly applied to a mucous membrane of eyes, nose, throat or ears. Such a formulation must satisfy the following two requirements: (1) the pH of the formulation is adjusted to be in the physiological pH range, i.e. pH 5 to 9, so as to reduce the irritation of the inflammatory mucous membrane of eyes, nose, throat and ears and (2) the product must be in the form of a clear aqueous solution. In other words, the solution must be filtered to remove bacteria, since the mucous membranes are liable to be infected with bacteria. For this purpose, lysozyme hydrochloride and dipotassium glycyrrhizinate must be dissolved to form a clear solution. Under these circumstances, the inventor made intensive investigations of the preparation of an aqueous solution containing lysozyme hydrochloride and dipotassium glycyrrhizinate which satisfies both of the above-mentioned requirements. However, the combination of lysozyme hydrochloride with dipotassium glycyrrhizinate immediately formed a precipitate in water to form a milky liquid and thus the preparation of the clear solution was impossible. Summary of the Invention After intensive investigations made for the purpose of solving the above-mentioned problems, the inventors have found that the problems can be solved by adjusting the pH of the solution to 5 to 9 and adding at least one of sodium chloride, potassium chloride, calcium chloride, magnesium chloride, sodium or potassium phosphate, sodium or potassium citrate and sodium or potassium carbonate. The present invention has been completed on the basis of this finding. Thus the present invention provides a solution containing lysozyme hydrochloride and dipotassium glycyrrhizinate characterized by having a pH adjusted to 5 to 9, and adding at least one of sodium chloride, potassium chloride, calcium chloride, magnesium chloride, sodium or potassium phosphate, sodium or potassium citrate and sodium or potassium carbonate. Subject matter of the present invention is an aqueous solution comprising lysozyme hydrochloride and dipotassium glycyrrhizinate according to claim 1. Preferred embodiments thereof are subject matter of claims 2 to 4. A further subject matter is the aqueous solution according to the present invention for use as a pharmaceutical or in cosmetic preparations according to claims 5 to 7. The pH of the solution of the present invention is limited to 5 to 9, preferably 6.5 to 8.0. For example, the pH of the eye drops is 7.0 to 7.5 in accordance with the pH of tears. This is a physiological condition necessitated for relieving the irritation of a mucous membrane when the solution is directly applied to the mucous membrane. Particularly when the mucous membrane to which the solution is to be applied is inflamed, its pH must be adjusted to be in the above-mentioned range, since the sensitivity of the membrane is increased by the inflammation. The solution of the present invention contains at least one of sodium chloride, potassium chloride, calcium chloride, magnesium chloride, sodium or potassium phosphate, sodium or potassium citrate and sodium or potassium carbonate. The sodium or potassium phosphate is preferably hydrogen- or dihydrogenphosphate, the sodium or potassium citrate is preferably hydrogen- or dihydrogencitrate, and the sodium or potassium carbonate is hydrogencarbonate. The amounts of the additives according to the present invention are not particularly limited, since they are determined depending on the amounts of lysozyme hydrochloride and dipotassium glycyrrhizinate. However, when lysozyme hydrochloride and dipotassium glycyrrhizinate are used in amounts of 0.1 wt. % and 0.25 wt. %, respectively, a preferred amount of sodium chloride is 0.5 to 1.5 wt. %, and that of sodium dihydrogenphosphate, sodium hydrogencarbonate, sodium hydrogenphosphate or potassium dihydrogenphosphate is 0.5 to 1.0 wt. %. The solution of the present invention is prepared by adding water to a mixture of lysozyme hydrochloride, dipotassium glycyrrhizinate and the additive(s) of the present invention, then adding sodium hydroxide thereto to adjust the pH of the solution to a given value and to obtain a solution, filtering the solution and, if necessary, sterilizing the filtrate. The solution of the present invention may further contain an antiseptic. An effect of the present invention consists in that a solution containing lysozyme hydrochloride and dipotassium glycyrrhizinate which is clear at a pH in a physiological pH range of 5 to 9 can be provided. Therefore, the solution of the present invention is used mainly as eye drops, nasal drops, ear drops, and other external drugs to be applied to the oral cavity and throat. However, the form of the solution is not limited to them and the solution in other forms can also be used. For example, the technique of the preparation of the solution containing lysozyme hydrochloride and dipotassium glycyrrhizinate according to the present invention as described above can be employed in the preparation of cosmetics. Thus, the aqueous solution of the present invention can be used also in the cosmetic field. [Examples] The following examples will further illustrate the present invention and the effects of the present invention. Example 1 900 mℓ of distilled water was added to a mixture of 1 g of lysozyme hydrochloride, 2.5 g of dipotassium glycyrrhizinate and 8 g of sodium chloride to obtain a solution. The pH of the solution was adjusted to 7.0 with 0.1 N sodium hydroxide. Distilled water was added thereto to make up a total volume of 1,000 mℓ. After filtration under sterile conditions, 5 mℓ of the solution was poured into a container sealed under sterile conditions to prepare eye drops. The products were clear. Example 2 900 mℓ of distilled water was added to a mixture of 1 g of lysozyme hydrochloride, 2.5 g of dipotassium glycyrrhizinate, 10 g of sodium dihydrogenphosphate and 1 g of methylparaben to obtain a solution. The pH of the solution was adjusted to 7.4 with 0.1 N sodium hydroxide. Distilled water was added thereto to make up a total volume of 1,000 mℓ. After filtration under sterile conditions, 2 mℓ of the solution was poured into a container sealed under sterile conditions to prepare eye drops. The products were clear. Example 3 Distilled water was added to a mixture of 5 g of lysozyme hydrochloride, 2.5 g of dipotassium glycyrrhizinate and 20 g of potassium dihydrogenphosphate to obtain a solution. The pH of the solution was adjusted to 7.8 with 0.1 N sodium hydroxide. Distilled water was added thereto to make up a total volume of 1,000 mℓ. After filtration under sterile conditions, 5 mℓ of the solution was poured into a container sealed under sterile conditions to prepare eye drops. The products were clear. Example 4 Distilled water was added to a mixture of 1 g of lysozyme hydrochloride, 2.5 g of dipotassium glycyrrhizinate and 20 g of potassium chloride to obtain a solution. The pH of the solution was adjusted to 6.2 with 0.1 N sodium hydroxide. Distilled water was added thereto to make up a total volume of 1,000 mℓ. After filtration under sterile conditions, 10 mℓ of the solution was poured into a container sealed to prepare nasal drops. The products were clear. Example 5 Distilled water was added to a mixture of 1 g of lysozyme hydrochloride, 2.5 g of dipotassium glycyrrhizinate and 20 g of sodium hydrogenphosphate to obtain a solution. Distilled water was added thereto to make up a total volume of 1,000 mℓ. The pH of the solution was 8.0. After filtration under sterile conditions, 20 mℓ of the solution poured into a container sealed under sterile conditions to prepare a drug to be applied to the throat. The products were clear. Example 6 Distilled water was added to a mixture of 5 g of lysozyme hydrochloride, 2.5 g of dipotassium glycyrrhizinate and 10 g of sodium hydrogencarbonate to obtain a solution. The pH of the solution was adjusted to 8.8 with 0.1 N sodium hydroxide. Distilled water was added thereto to make up a total volume of 1,000 mℓ. After filtration under sterile conditions, 5 mℓ of the solution was poured into a container sealed under sterile conditions to prepare ear drops. The products were clear. Example 7 Distilled water was added to a mixture of 1 g of lysozyme hydrochloride, 2.5 g of dipotassium glycyrrhizinate and 20 g of calcium chloride to obtain a solution. The pH of the solution was adjusted to 7.3 with 0.1 N sodium hydroxide. Distilled water was added thereto to make up a total volume of 1,000 mℓ. After filtration under sterile conditions, 5 mℓ of the solution was poured into a container sealed under sterile conditions to prepare eye drops. The products were clear. Example 8 Distilled water was added to a mixture of 0.1 g of lysozyme hydrochloride, 0.5 g of dipotassium glycyrrhizinate and 8 g of sodium chloride to obtain a solution. The pH of the solution was adjusted to 7.5 with 0.1 N sodium hydroxide. Then 40 g of glycerol, 20 g of sorbitol, 2 g of HCO-50 [polyoxyethylene hardened castor oil (50 mol of ethylene oxide added)] and a suitable amount of a flavor were added to the solution. The pH of the solution was adjusted to 7.0 with a solution of 1 g of methylparaben in 80 mℓ of ethanol. Distilled water was added thereto to make up a total volume of 1,000 mℓ. It was filtered to obtain a lotion. This lotion was clear. Example 9 Distilled water was added to a mixture of 1 g of lysozyme hydrochloride, 2.5 g of dipotassium glycyrrhizinate, 3 g of sodium hydrogenphosphate and 8 g of sodium chloride to obtain a solution. The pH of the solution was adjusted to 6.8 with 1 N sodium hydroxide. Distilled water was added thereto to make up a total volume of 1,000 mℓ. This solution was clear. Example 10 Distilled water was added to a mixture of 1 g of lysozyme hydrochloride, 2.5 g of dipotassium glycyrrhizinate, 1 g of potassium chloride and 8 g of sodium chloride to obtain a solution. The pH of the solution was adjusted to 7.2 with 1 N sodium hydroxide. Distilled water was added thereto to make up a total volume of 1,000 mℓ. This solution was clear. Example 11 Distilled water was added to a mixture of 1 g of lysozyme hydrochloride, 2.5 g of dipotassium glycyrrhizinate, 0.3 g of calcium chloride and 8.5 g of sodium chloride to obtain a solution. The pH of the solution was adjusted to 7.1 with 1 N sodium hydroxide. Distilled water was added thereto to make up a total volume 1,000 mℓ. This solution was clear. Example 12 Distilled water was added to a mixture of 1 g of lysozyme hydrochloride, 2.5 g of dipotassium glycyrrhizinate, 70 mg of potassium dihydrogenphosphate and 8.5 g of sodium chloride to obtain a solution. The pH of the solution was adjusted to 7.0 with 1 N sodium hydroxide. Distilled water was added thereto to make up a total volume 1,000 mℓ. This solution was clear. Example 13 Distilled water was added to a mixture of 1 g of lysozyme hydrochloride, 2.5 g of dipotassium glycyrrhizinate, 2 g of sodium hydrogencarbonate and 7 g of sodium chloride to obtain a solution. The pH of the solution was adjusted to 7.0 with 1 N sodium hydroxide. Distilled water was added thereto to make up a total volume of 1,000 mℓ. After filtration under sterile conditions, 5 mℓ of the solution was poured into each of sealed containers under sterile conditions to prepare dye drops. The products were clear. Example 14 Distilled water was added to a mixture of 1 g of lysozyme hydrochloride, 2.5 g of dipotassium glycyrrhizinate, 8 g of sodium chloride and 2 g of sodium dihydrogenphosphate to obtain a solution. The pH of the solution was adjusted to 7.3 with 1 N sodium hydroxide. Distilled water was added thereto to make up a total volume of 1,000 mℓ. This solution was clear. Example 15 Distilled water was added to a mixture of 1 g of lysozyme hydrochloride, 2.5 g of dipotassium glycyrrhizinate, 4 g of sodium dihydrogenphosphate and 6 g of sodium chloride to obtain a solution. The pH of the solution was adjusted to 6.8 with 1 N sodium hydroxide. Distilled water was added thereto to make up a total volume of 1,000 mℓ. This solution was clear. Example 16 Distilled water was added to a mixture of 1 g of lysozyme hydrochloride, 2.5 g of dipotassium glycyrrhizinate, 3 g of sodium citrate and 8 g of sodium chloride to obtain a solution. The pH of the solution was adjusted to 7.0 with 1 N sodium hydroxide. Distilled water was added thereto to make up a total volume of 1,000 mℓ. This solution was clear. Example 17 Distilled water was added to a mixture of 1 g of lysozyme hydrochloride, 2.5 g of dipotassium glycyrrhizinate, 6 g of sodium chloride and 8.3 g of sodium hydrogenphosphate to obtain a solution. The pH of the solution was adjusted to 7.0. Distilled water was added thereto to make up a total volume of 1,000 mℓ. This solution was clear. Example 18 Distilled water was added to a mixture of 1 g of lysozyme hydrochloride, 2.5 g of dipotassium glycyrrhizinate, 8 g of sodium chloride, 70 mg of sodium dihydrogenphosphate, 200 mg of potassium chloride and 200 mg of calcium chloride to obtain a solution. The pH of the solution was adjusted to 7.0 with 1 N sodium hydroxide. Distilled water was added thereto to make up a total volume of 1,000 mℓ. This solution was clear. Example 19 Distilled water was added to a mixture of 1 g of lysozyme hydrochloride, 2.5 g of dipotassium glycyrrhizinate, 7 g of sodium chloride, 1.0 g of potassium chloride and 1.0 g of sodium dihydrogenphosphate to obtain a solution. The pH of the solution was adjusted to 7.0. Distilled water was added thereto to make up a total volume of 1,000 mℓ. This solution was clear. Example 20 Distilled water was added to a mixture of 1 g of lysozyme hydrochloride, 2.5 g of dipotassium glycyrrhizinate, 1 g of potassium chloride, 5.5 g of sodium chloride, 10 g of sodium hydrogenphosphate and 2 g of sodium dihydrogenphosphate to obtain a solution. The pH of the solution was adjusted to 7.0. Distilled water was added thereto to make up a total volume of 1,000 mℓ. This solution was clear. Example 21 Distilled water was added to a mixture of 1 g of lysozyme hydrochloride, 2.5 g of dipotassium glycyrrhizinate, 8 g of sodium chloride, 70 mg of potassium dihydrogenphosphate and 0.2 g of potassium chloride to obtain a solution. The pH of the solution was adjusted to 7.0. Distilled water was added thereto to make up a total volume of 1,000 mℓ. This solution was clear. Example 22 Distilled water was added to a mixture of 1 g of lysozyme hydrochloride, 2.5 g of dipotassium glycyrrhizinate, 8 g of sodium chloride, 50 mg of potassium dihydrogenphosphate and 0.2 g of calcium chloride to obtain a solution. The pH of the solution was adjusted to 7.0. Distilled water was added thereto to make up a total volume of 1,000 mℓ. This solution was clear. Example 23 Distilled water was added to a mixture of 1 g of lysozyme hydrochloride, 2.5 g of dipotassium glycyrrhizinate, 8.5 g of sodium chloride, 2 g of potassium chloride and 0.2 g of calcium chloride to obtain a solution. The pH of the solution was adjusted to 7.0. Distilled water was added thereto to make up a total volume of 1,000 mℓ. This solution was clear. Example 24 Distilled water was added to a mixture of 1 g of lysozyme hydrochloride, 2.5 g of dipotassium glycyrrhizinate, 5.5 g of sodium chloride, 0.2 g of potassium chloride and 0.15 g of sodium hydrogenphosphate to obtain a solution. The pH of the solution was adjusted to 7.0. Distilled water was added thereto to make up a total volume of 1,000 mℓ. This solution was clear. Example 25 900 mℓ of distilled water was added to a mixture of 1 g of lysozyme hydrochloride, 2.5 g of dipotassium glycyrrhizinate and 15 g of sodium chloride to obtain a solution. The pH of the solution was adjusted to 5.0 with 0.1 N sodium hydroxide. Distilled water was added thereto to make up a total volume of 1,000 mℓ. This solution was clear. Example 26 900 mℓ of distilled water was added to a mixture of 1 g of lysozyme hydrochloride, 2.5 g of dipotassium glycyrrhizinate and 5 g of sodium hydrogencarbonate to obtain a solution. The pH of the solution was adjusted to 5.0 with 0.1 N hydrochloric acid. Distilled water was added thereto to make up a total volume of 1,000 mℓ. This solution was clear. Example 27 Distilled water was added to a mixture of 1 g of lysozyme hydrochloride, 2.5 g of dipotassium glycyrrhizinate, 0.3 g of magnesium chloride and 9 g of sodium chloride to obtain a solution. The pH of the solution was adjusted to 7.0 with 1 N sodium hydroxide. Distilled water was added thereto to make up a total volume of 1,000 mℓ. This solution was clear.

Claims: Claims for the following Contracting States : AT, BE, CH, DE, FR, GB, IT, LI, LU, NL, SE An aqueous solution which comprises lysozyme hydrochloride in an amount of or below 0.5 % by weight, dipotassium glycyrrhizinate in an amount of below 0.3 96 by weight, at least one member selected from sodium chloride, potassium chloride, calcium chloride, magnesium chloride, sodium phosphate, potassium phosphate, sodium citrate, potassium citrate, sodium carbonate and potassium carbonate and water and has a pH adjusted to 5 to 9. The aqueous solution as claimed in Claim 1, which comprises 0.1 to 0.5 percent by weight of lysozyme hydrochloride and 0.05 to 0.25 percent by weight of dipotassium glycyrrhizinate. The aqueous solution as claimed in Claim 1, in which the phosphate is hydrogenphosphate or dihydrogenphosphate, the citrate is hydrogencitrate or dihydrogencitrate and the carbonate is hydrogencarbonate. The aqueous solution as claimed in Claim 1, which has a pH of 7.0 to 7.5. The aqueous solution according to claims 1 to 4 for use as a pharmaceutical. The aqueous solution according to claim 5 for the preparation of eye drops, nasal drops, ear drops and external compositions for the treatment of the oral cavity and throat. The aqueous solution according to claims 1 to 4 for use in cosmetic preparations. Claims for the following Contracting States : GR, ES A process for the preparation of an aqueous solution containing lysozyme hydrochloride and dipotassium glycyrrhizinate, characterized by dissolving lysozyme hydrochloride, dipotassium glycyrrhizinate, at least one member selected from sodium chloride, potassium chloride, calcium chloride, magnesium chloride, sodium phosphate, potassium phosphate and sodium citrate, potassium citrate, sodium carbonate and potassium carbonate in water and adjusting the solution to a pH of 5 to 9, and in that lysozyme hydrochloride is added in an amount of or below 0.5 % by weight, and dipotassium glycyrrhizinate is added in an amount of below 0.3 % by weight. The process according to claim 1 characterized in that lysozyme hydrochloride is added in an amount from 0.1 to 0.5 % by weight, and dipotassium glycyrrhizinate is added in an amount from 0.05 to 0.25 % by weight. The process of claims 1 or 2 characterized in that the phosphate is selected from hydrogenphosphate or dihydrogenphosphate, the citrate is selected from hydrogencitrate or dihydrogencitrate, and the carbonate is selected from hydrogencarbonate. The process of claims 1 to 3 characterized in that the aqueous solution has a pH of 7.0 to 7.5. The process according to any of claims 1 to 4, characterized in that the aqueous solution is formulated to eye drops, nasal drops, ear drops or an external composition for the treatment of the oral cavity and throat. The use of an aqueous solution prepared according to any of claims 1 to 4 for the preparation of a medicament useful against bacterial infections.

Inventors: KOGURE, YOSHIO
Applicants: EISAI CO., LTD.
Application year: 1987
Main classification: A61K37
IPC codes: A61P27, A61K47, A61P37, A61K8, A61K9, A61K31, A61K38, A61P29, A61P1