JPS649295B2 - - Google Patents

Description

[Detailed description of the invention]

The present invention relates to a novel topical preparation, and more particularly to a topical preparation containing urokinase as an active ingredient. Urokinase is produced by extraction from human urine or by culturing kidney tissue, and has a molecular weight of approximately 3 to 3.
It is a 50,000 protein enzyme and has been widely used clinically as a thrombolytic agent. In addition, in recent years, it has been reported that intravenous administration of this urokinase is effective in treating skin diseases based on fibrin deposition, such as systemic scleroderma and Juering's dermatitis herpetiformis ("Clinical and Research"). Volume 49, No. 2962~
2964, 1972). Therefore, the effect of urokinase is to activate plasminogen activator in blood vessels or tissues, thereby enhancing fibrinolytic activity, removing and suppressing fibrin deposition and tissue fibrosis. It is said that this is something that can be demonstrated through this process. Therefore, from the viewpoint of the mechanism of action of urokinase and its stability, it has been conventionally said that urokinase is preferably administered intravenously, and other administration methods have not been investigated. However, with intravenous administration of urokinase, when treating peripheral venous blood clots or local skin diseases, the amount of urokinase that reaches the local thrombus or skin lesion site is extremely small, so a large amount of urokinase is administered. Along with the need to
It has the disadvantage that it imposes a physical burden on patients because it requires long-term administration. Furthermore, urokinase is produced through a complicated extraction and purification process, and is an enzyme whose activity tends to decrease, making it extremely expensive. Therefore, the above-mentioned large-scale administration poses a major bottleneck from an economic standpoint. Under these circumstances, the present inventors have proposed an effective method for administering urokinase, especially considering that transdermal administration of urokinase is difficult since it is a high-molecular-weight protein and its activity is likely to decrease. As a result of various studies on topical urokinase, which had not been studied at all,
Contrary to the above predictions, it was discovered that urokinase, when made into an oil-in-water ointment, is effectively absorbed through the skin and has excellent effects in treating local skin diseases and peripheral blood clots, and the present invention was completed. did. Therefore, the present invention provides urokinase 100~
The present invention provides a topical urokinase preparation consisting of an oil-in-water ointment containing 100,000 IU/g. The external preparation for urokinase of the present invention is produced by blending a predetermined amount of urokinase in a conventional manner using a commonly used oil-in-water ointment base. The amount of urokinase blended into the external preparation can be increased or decreased as appropriate, but it is preferably blended between 100 and 100,000 IU/g. Next, the results of tests on transdermal absorption and stability of the urokinase topical preparation of the present invention are as follows. (1) Percutaneous absorption test The abdominal skin of a hairless mouse (male, 5 weeks old) was cut out, placed in a glass tube (cross section 1 cm ² ), and various urokinase-containing ointments were applied from inside the glass tube.
This was soaked in 10 ml of physiological saline. Note that the physiological saline solution was kept stirring during the test. 1 hour, 3 hours and 6
After a period of time, 0.5 ml of physiological saline was collected, and urokinase activity and fibrinolytic activity (plasmin activity) based on skin tissue activator release in the solution were measured. The ointment shown in the table was used as the measurement sample.

[Table] Base ^** : Ointment base of Example 2 In addition, synthetic substrate S-2444 was used to measure the amount of urokinase.
(Pyro Glu-Gly-Arg-pNA) and S-2251 (H・D-Val-Leu-Lys-
pNA) was used. Urokinase amount measurement Sample 0.5ml and synthetic substrate S-2444 (manufactured by Kabi Co., Ltd.)
0.5 ml of the 200 μg/ml solution was mixed and incubated at 37°C. 50% acetic acid after 2 and 4 hours.
Hydrolysis was stopped by adding 1.5 ml, and the absorbance at 405 nm was measured and calculated as the amount of urokinase. The calibration value is urokinase 10IU/ml.
5IU/ml and 2.5IU/ml solutions were prepared by processing in the same manner as the above samples. The calibration values are shown in the table, and the measurement results are shown in the table.

【table】

[Table] Plasmin amount measurement Sample 0.5ml and synthetic substrate S-2251 (manufactured by Kabi Co., Ltd.)
Using 0.5 ml of the 200 μg/ml solution, the amount of plasmin was determined by processing in the same manner as described below. The calibration values are 10×10 ^-3 cu/ml, 5×10 ^-3 cu/ml and plasmin.
A sample of 2.5×10 ^-3 cu/ml was prepared in the same manner. The calibration values are shown in Table V, and the measurement results are shown in Table V.

【table】

[Table] As is clear from the above experiment, the plasmin activity based on the release of skin tissue activator is almost the same in both cases, including the base agent and urokinase.
This is because the substrate S in the experiment is extremely small.
It can be seen that it is not involved in the hydrolysis of -2444. Therefore, the difference in the degree of hydrolysis between the base alone and the base containing urokinase in the experiment is due to urokinase, which means that urokinase is absorbed transdermally. (2) Stability test Urokinase ointments prepared according to Examples 1 and 2 were stored at room temperature and in the refrigerator, and the urokinase activity was measured after 3 and 14 days. The results are shown in the table.

[Table] Next, the urokinase activity was measured when the ointment was stored at 5°C for 4 months. The results are shown in the table.

[Table] As is clear from this test, the urokinase topical preparation of the present invention is stable and can be stored for a long period of time if refrigerated, and can be stored for a considerable period of time even at room temperature. As mentioned above, the topical preparation of urokinase of the present invention can raise the concentration of urokinase at the lesion site higher than that by intravenous injection by the simple administration method of applying it to the skin of the local lesion site. Urokinase acts directly and quickly on the affected area and can effectively cure the disease, and has few side effects, making it extremely excellent medically. Therefore, the topical urokinase preparation of the present invention is useful for diseases treated by intravenous injection such as peripheral varicose veins and thrombophlebitis, as well as eczema, prurigo, erythema, and purpura;
Various types of vasculitis such as cutaneous periarteritis nodosa, persistent erythema erectus, temporal arteritis, Mondor's syndrome, and idiopathic gangrene; blood circulation disorders, gangrenous diseases; burns, chemical injuries, frostbite,
Physical and chemical disorders such as frostbite, electric shock injury, and radiation damage; Bullosa, pustulosis, and keratoderma; Inflammatory keratosis such as psoriasis, psoriasis, and progressive palmar keratoderma; Skin sclerosis,
It can be used to treat dermatomyositis, lupus erythematosus, neonatal subcutaneous fat necrosis, epithelial tumors, and pilosebaceous diseases such as acne vulgaris. It can also be used for purposes such as promoting healing of Bazin's erythema induration and tuberculous phlebitis, enhancing the action of anticancer agents for skin cancer, enhancing the action of steroids, and preventing side effects. As mentioned above, the method of administering the topical preparation of urokinase of the present invention includes not only the method of applying it directly to the affected area or the skin of the affected area, but also the occlusive bandage therapy in which it is sealed with tape, cellophane, etc. Examples and clinical examples will be further described below. Example 1 Urokinase 1.0g Stearic acid 9.0g Sodium stearate 1.5g Setanol 2.5g Isopropyl myristate 5.0g Surfactant 8.9g Preservative 0.2g Purified water Total amount 100.0g An O/W ointment was prepared using the above ingredients. Example 2 Urokinase 1.0g Vaseline 4.0g Liquid paraffin 6.0g Setanol 3.0g Stearyl alcohol 3.0g Isopropyl myristate 4.0g Surfactant 3.5g Purified water Total amount 100.0g An O/W ointment was prepared using the above ingredients. Example 3 Urokinase 1.0g Stearic acid 25.0g Setanol 1.0g Lanolin 1.0g Glycerin 5.0g Triethanolamine 3.0g Purified water Total amount 100.0g An O/W ointment was prepared from the above ingredients. The following clinical examples all show the results of using the external ointment (containing urokinase 10,000 IU/g) produced in Example 1 on burn cases. Clinical case 1 Patient: Minami 〇ko, 1 year and 8 months old Girl First examination: July 2, 1978 Chief complaint: Burns on the left side and both arms History of current illness: Injured in boiling water around 10 a.m. on June 30; I rinsed it off with cold water for a few minutes and received ointment treatment at a certain clinic. Current symptoms: There are lesions the size of the palm of a child's hand in the left vulva and right cubital fossa, and lesions covering almost the entire left arm. Although the blister contents have already been removed, almost the entire surface is covered with the blister membrane. Diagnosis: Second-degree superficial burn Treatment and course: Based on the above diagnosis, after removing the membrane, a sufficient amount of urokinase-containing ointment was applied locally and bandaged, and the bandage was changed every 48 hours. On July 4th, the local symptoms had significantly improved, and there was no exudate, moss, or swelling, and erythematous changes were predominant, so from July 7th onwards, I started using urokinase-containing ointment. I stopped using it and simply applied zinc borate ointment. Side effects: None observed. Effects of UK-containing ointment: It quickly dries the affected area and is considered effective. Clinical Case 2 Patient: Manabu Miya, 1 year old, boy First examination: June 27, 1976 Chief complaint: Burns on the right wrist History of current illness: Injury caused by boiling water on June 26, left untreated. Current symptoms: There is a pigeon-egg-sized bean on his right wrist. Diagnosis: Second-degree superficial burn treatment and course: Based on the above diagnosis, a sufficient amount of urokinase-containing ointment was applied locally and a bandage was applied, which was changed once a day. By July 3rd, he had completely recovered. Side effects: None observed. Effect of urokinase-containing ointment: Complete recovery was achieved promptly. Therefore, it is recognized as extremely effective. Clinical Case 3 Patient: Toru Sako, 1 year and 6 months old. First examination: July 16, 1971. Chief complaint: Burn injury on left wrist. History of current illness: 30 minutes before consultation, the injury was caused by boiling water, and the injury was cooled with tap water for about 5 minutes. . Current symptoms: The area distal to the left wrist is flushed, swollen, and appears to be painful. The lesions are most obvious in the wrist joints, where blisters and ruptured blisters are observed. Diagnosis: Second-degree superficial burn treatment and course: Based on the above diagnosis, a sufficient amount of urokinase-containing ointment was applied to the affected area, and then bandaged and changed every 48 hours. By July 18, the local redness, swelling, and exudate had all become very slight, and although no topical antibiotics were used, no changes were observed that would suggest secondary infection. By July 20th, only mild erythema remained locally. Side effects: None observed. Effects of urokinase-containing ointment: Prompt improvement of local dryness and inflammatory symptoms. It is recognized as extremely effective.

Claims (1)

[Claims] 1. Urokinase external preparation consisting of an oil-in-water ointment containing 100 to 100,000 IU/g of urokinase.