JPH0577656B2 - - Google Patents

Description

[Detailed description of the invention]

【0001】

TECHNICAL FIELD The present invention relates to a therapeutic agent for peripheral vascular disorders, and particularly to a therapeutic agent for peripheral vascular disorders containing kallidinogenase as an active ingredient.

【0002】

[Prior art and its problems] Kallidinogenase is an enzyme produced in the pancreas and excreted in the urine.
It is thought that kallidin is formed from kininogen and acts on the antihypertensive system by directly or indirectly affecting water electrolyte metabolism in the kidney, and is said to be effective in various circulatory system disorders such as hypertension, peripheral vascular disorders,
It is useful as a therapeutic agent for angina pectoris, etc. Conventionally, kallidinogenase extracted mainly from pig pancreas has been administered orally or by injection to treat the above-mentioned circulatory system disorders, but when kallidinogenase is administered orally or by injection to treat peripheral vascular disorders, Since the amount of kallidinogenase that is reached is extremely small, large amounts of administration over a long period of time are required, and as a result, side effects may occur. From this safety standpoint, kallidinogenase, which is derived from human urine instead of pig pancreas, has recently attracted attention. However, even if kallidinogenase is derived from human urine, there are concerns that it may cause side effects when administered orally in large quantities over a long period of time for the purpose of treating peripheral vascular disorders.

【0003】

OBJECTS OF THE INVENTION Accordingly, an object of the present invention is to provide a kallidinogenase preparation for treating peripheral vascular disorders with no or extremely low risk of side effects.

【0004】

[Means and effects for solving the problem and achieving the object] The present inventors have conducted intensive studies on the administration methods, therapeutic effects, etc. of kallidinogenase of various origins, and have found that kallidinogenase has percutaneous absorption ability. The present invention was completed by discovering that

[0005] Therefore, the peripheral vascular disorder therapeutic agent according to the present invention is characterized by being an external preparation containing kallidinogenase as an active ingredient.

[0006] That is, unlike conventional oral preparations and injection preparations, the therapeutic agent for peripheral vascular disease according to the present invention is an external preparation, and since it is administered locally, kallidinogenase can be effectively delivered to the affected area. ,
Therefore, the possibility of side effects occurring can be significantly reduced. Exemplary dosage forms of the therapeutic agent for peripheral vascular disorders according to the present invention include external creams, external gel ointments, external solutions, and poultices. Incidentally, citrate may be added for the purpose of stabilizing the main agent, kallidinogenase. The amount of this citrate to be blended is preferably 0.1-10% by weight as sodium citrate.

[0007] The therapeutic agent for peripheral vascular disease according to the present invention is useful not only for the treatment of pure peripheral vascular disease, but also for the treatment of various skin diseases thought to be caused by quasi-peripheral vascular disease, such as scleroderma, Raynaud's syndrome, and venous disease. It is. Since the therapeutic agent for peripheral vascular disease according to the present invention is an external preparation, the possibility of side effects is extremely low. Therefore, the main component, kallidinogenase, may be derived from animals other than humans, and its stabilization It can also be a kallidinogenase modified with a derivative such as polyethylene glycol. When formulating the therapeutic agent for peripheral vascular disorders according to the present invention, conventional pharmaceutical carriers such as vaseline, glycerin, and cyclodextrin, and conventional absorption aids such as triethanolamine and Tween (trademark) may be added. can. The amount of kallidinogenase to be blended is determined depending on the type and severity of the disease, but it is usually within the range of 1-1000 international kallidinogenase units (IU)/g [Kallidinogenase activity is indicated by "KU", "KE" ”, etc., but later “IU” or “International Unit” was used.
“IU”)]

[0008]

[Production Examples, etc.] Next, the present invention will be explained in more detail and concretely using reference examples, production examples, and test examples.

Reference Example 1 (Production method of modified kallidinogenase) 2-o-methoxypolyethylene glycol-
200 mg of 4,6-dichloro-S-triazine and 25 mg of kallidinogenase were reacted in a 0.1 M borate buffer under low temperature conditions for 2 hours, and then ultrafiltered.
Polyethylene glycol-modified kallidinogenase was obtained by freeze-drying.

[0010] Production Example 1 (Cream for external use) A cream for external use was prepared by mixing the following ingredients by a conventional method. Porcine pancreatic kallidinogenase 0.5 (g) (1.1 million biological units) Sodium benzoate 0.5 Diethyl sebatate 10.0 Sodium polyoxyethylene oil ether phosphate 6.0 Vaseline balance Total 100.0

[0011] Production Example 2 (Gel ointment for external use) A gel ointment for external use was prepared by mixing the following components by a conventional method. Kallidinogenase 1 (g) (2.2 million biological units) Sodium citrate 3 Stearic acid 25 Setanol 1 Lanolin 1 Glycerin 5 Triethanolamine 3 Purified water remainder Total 100

Production Example 3 (Liquid for external use) A liquid for external use was prepared by mixing the following components in a conventional manner. Kallidinogenase 1 (g) (2.2 million product biological units) Cyclodextrin 2 Sodium citrate 1 Glycerin 3 Purified water remainder Total 100

[0013] Production example 4 (topical patch) 1.5 g of sodium polyacrylate and 0.6 g of Carbopol 90 (trademark) are mixed, the following ingredients are added to this, and purified water is added to make the total amount. A poultice substrate was prepared by adding 100 g of the powder and then further mixing. Gelatin 6 (g) Tween 80 (trademark) 3 Glycerin 30 Callidinogenase 1 (2.2 million biological units) The above poultice substrate is coated onto a poultice carrier to a thickness of 3 mm or less, and each sheet contains kallidinogenase. A patch containing 500,000 biological units was obtained.

[0014] Test Example 1 (Transdermal Absorption Test) The present applicant has developed a reagent for measuring urine kallidinogenase, and this reagent is commercially available as "Naftest-UK (trade mark)", so the measurement method using this reagent is Similarly, the transdermal absorption potential of the formulation according to the present invention was investigated. a) Specimen The abdominal skin of a male rat was cut out, a patch according to Formulation Example 4 was applied, and after 60 minutes, 1 cm ² of this skin was immersed in 10 ml of physiological saline and stirred for 120 minutes to prepare a specimen. b) Reagents) Buffer 100mM phosphate buffer (PH7.0) prepared using 0.03% sodium laurate solution. ) Add purified water to 5.56 mg of substrate prolylphenylalanylarginine naphthyl ester (PPANE),
Prepared by adjusting the total volume to 3 ml. ) Color forming agent Prepared by adding purified water to 60 mg of Fastret ITR salt to make a total volume of 6 ml. ) Antitropin 4-methyl-1-[N-(3-methyl-1,2,
10 ^-4 g of 3,4-tetrahydro-8-quinolinesulfonyl)-L-arginyl]-2-piperidinecarboxylic acid (MD-805) per ml of the above buffer.
Prepared to contain. ) Reaction terminator: Use pure glacial acetic acid. c) Test method for measuring kallidinogenase Collect 0.1 ml each of the above specimen, buffer solution, substrate, and antitropin agent, stir and mix, and then
Shake for 30 minutes at °C. Next, 0.1 ml of the above color former is added, stirred, and left at 37°C for 5 minutes, and then 1.0 ml of glacial acetic acid is added to stop the azo dye formation reaction. The absorbance of the resulting solution was measured at the wavelength within 1 hour of the addition of glacial acetic acid.
Measured at 475nm. On the other hand, a standard calibration curve plotting the relationship between absorbance and kallidinogenase concentration was prepared in advance by using a pure product of kallidinogenase instead of the above sample, setting the kallidinogenase concentration to various values, and carrying out the same method as above. Oita. The absorbance value in this standard calibration curve is compared with the absorbance value measured for the above specimen to determine the kallidinogenase concentration for the specimen. d) Test results The measured concentration of kallidinogenase in the sample is
The amount was 0.035 NU/ml (NU: α-naphthol unit), which indicates that kallidinogenase can be absorbed transdermally.

Test Example 2 (Acute Toxicity) Kallidinogenase is an enzyme (glycoprotein) produced by living organisms, and its toxicity varies depending on the species of the animal of origin, and physicochemical differences are also observed depending on the animal of origin. Therefore, it is difficult to confirm the safety of clinical use from the results of animal experiments, but the acute toxicity when kallidinogenase derived from human urine is administered orally or intravenously to rats is as follows. Considering that the peripheral vascular disorder therapeutic agent is an external preparation, it can be said that the toxicity of the preparation according to the present invention is extremely low.

【0016】 ■■■ Turtle shell [0052] ■■■

[0017] Test Example 3 (Percutaneous Absorption Test) Confirming the therapeutic effect of kallidinogenase on peripheral vascular disorders through animal experiments is difficult because it is difficult to create diseased animals and perform suction and excretion tests. We investigated the transdermal absorption effect of nitroglycerin, which is thought to be due to its action. a) Test method The backs of male Wistar rats weighing 245-410 g were removed, and 50% of nitroglycerin ointment was applied to a lint cloth.
A transdermal absorption test of nitroglycerin was performed by applying a mixture of 1 mg/mouse as nitroglycerin and 50 mg of the gel ointment according to Production Example 2 (11 IU/mouse as kallidinogenase). On the other hand, as a control test, a lint cloth coated with only nitroglycerin ointment was applied to the hair-removed backs of rats to conduct a transdermal absorption test of nitroglycerin. In addition, absorption is determined by the concentration of nitroglycerin in plasma.
It was determined by measuring according to the method of Pete SK et al. ["J.Pharm.Soc." Vol. 67, p. 584 (1978)]. b) Test results Both the test group and the control group reached the maximum concentration after 15 minutes, and the nitroglycerin concentration values were 14 mg/ml in the test group and 11 mg/ml in the control group.
This indicates that kallidinogenase enhances the transdermal absorption ability of nitroglycerin, in other words, kallidinogenase itself has the transdermal absorption ability.

[0018] Test Example 4 (Usage Test) Subjects were those who felt numbness in their hands and fingers during cold weather and those who were exhibiting keratin symptoms and who were recognized as suffering from semi-peripheral vascular disease. A topical cream was given to the patients and used as a hand cream for 10 days.Afterwards, the symptoms were classified into four categories: () improved, () I think it's getting better, () Same as before, and) () Worse. When a questionnaire survey was conducted regarding this, all of the respondents answered that they had "improved" or "I think they are improving," confirming the effectiveness of the therapeutic agent according to the present invention.

【0019】

[Effect of the invention] Conventionally, kallidinogenase preparations administered for the purpose of treating peripheral vascular disorders have been administered in the form of oral preparations or injections, and therefore large doses have been required to ensure that a sufficient amount of the drug reaches the affected area. There is a need for this, and since treatment of this type of disease requires a long period of time, there is a possibility of side effects occurring. Therefore, the use of human urine kallidinogenase has been considered in place of porcine pancreatic kallidinogenase, which is more likely to cause side effects, but there is still concern about the possibility of side effects caused by large amounts and long-term use. In contrast, the therapeutic agent for peripheral vascular disease according to the present invention contains kallidinogenase as an active ingredient, but it is an external preparation, and since kallidinogenase is absorbed transdermally and acts on the affected area, it cannot be used in conventional oral preparations or injections. The amount reaching the affected area is increased and the possibility of side effects is lower when compared to

Claims (1)

[Scope of Claims] [1] A therapeutic agent for peripheral vascular disease, which is an external preparation containing kallidinogenase as an active ingredient. [2] The dosage form is external cream, external gel ointment,
The therapeutic agent for peripheral vascular disease according to claim 1, which is selected from external liquid preparations and topical tablets. [3] A therapeutic agent for peripheral vascular disorders, which is an external preparation containing kallidinogenase as an active ingredient, and is characterized by containing citrate. [4] Sodium citrate as citrate
characterized by containing 0.1-10% by weight,
The peripheral vascular disorder therapeutic agent according to claim 3.