KR930002011B1 - Antihypertensive and antihpotensive preparations - Google Patents

Abstract

No content.

Description

[Name of invention]

Blood pressure regulators

Detailed description of the invention

The present invention relates to a blood pressure regulator or an antihypertensive blood pressure agent. More specifically, the antihypertensive or antihypertensive blood pressure, which has an excellent antihypertensive action and / or antihypertensive action and stability, and is composed of an active ingredient in a pharmacologically acceptable worm dry powder by a novel and advanced improved formulation. It's about the first.

In recent years, many patients with hypertension or hypotension, regardless of old age. For the treatment of these patients, hypertensives are administered to blood pressure lowering agents, and hypotension patients are not necessarily required, but blood pressure increasing agents (or nicknames, boosters) are often administered.

In particular, with the increase of hypertension patients, a number of various hypertension therapeutic agents (or blood pressure lowering agents) have been developed and used. In patients with hypertension, if the patient stops taking the blood pressure lowering drug after starting the blood pressure lowering, the bed may be congested and become heavy. For this reason, there are many cases where you must take medicine for a long time. Patients with hypotension often receive long-term administration when using drugs. Therefore, the drug which does not have a side effect is desired.

For example, hydralazine, which is known as a blood pressure lowering agent, has a peripheral blood vessel dilation action and shows excellent blood pressure lowering action, but since it causes tachycardia as a side effect, it is used in combination with β-blocker. In general, drugs that have no side effects are particularly desired because of the long-term administration of the blood pressure lowering agent and the blood pressure raising agent.

On the other hand, earthworms have been used as medicines since ancient times, mainly called the Guin (지) and Jiryong (地 章) in the Oriental Empire. The pharmacological and medicinal effects of the earthworm reported in the conventional literature are shown below.

① "Earth and life" Ofuchi Shinryuzer October 30, 1947 (22 years of fire extinguishing), Maki West outbreak, pages 223-226, and "Relayed earthworm" The earthworm has a pharmacological action on the 160-163 pages of earthworms, the reducing agent of the stones in the bladder and the excretion agent out of the body, the treatment of jaundice, the delivery agent, the tonic, the mother medicine, the anti-corrosive agent and the antipyretic agent. On the other hand, it is reported that as an earthworm poison, one invades the nervous system and the other has the destruction of red blood cells, that is, hemolysis.

(2) Pharmacopoeia of the People's Republic of China Pharmacopoeia Committee of the People's Republic of China, 1977 edition, Part 1 pages 197 to 198.

By convention, there are two kinds of Jiryong products. One is Lumbriscus Kwangtungesis, which splits the abdomen, washes off viscera and mud sand, and dries it in the sun, shade or cold. The other land dragon (Lumbricus Nativus) was killed by putting it in vegetation ash and then removing the ash and drying it in the sun, shade, or low temperatures. Efficacy and application of these two kinds of earth dragons are fever, calming winds, calming panting by breathing, through the route (good circulation), winds and cramps due to high fever, paralysis and limbs Involuntary, swelling and swelling of the joints, incomplete urination, panting of the bronchus, hypertension, and reported to be used at 4.5 to 9 g / day.

③ `` Our Herbal Medicine Series 3, Jilong, Squid Bones, Scientific Research in China '' issued on October 30, 1978, Matsuura Pharmaceutical Co., Ltd. Is reporting.

(4) Digestion of Natural Drug Dictionary On April 15, 61, Hirokawa Bookstore issued, page 215, reports that Jiryong is used for antipyretics, analgesics, diuretics and antidote.

⑤ Tanaka Mamoru, Japan Hokkaido Medical Journal, Vol. 24, pp. 18 to 24 (1949), obtained by removing nitolol from old dry matter from boiling water and adding ethyl alcohol to the concentrate of the extract. (Lumbrofebrin) was dissolved in Ringer's seed solution and intravenous injection of this solution into cats under anesthesia resulted in a sudden drop in blood pressure, and reported to promote blood coagulation in proportion to shock.

⑥ Kenjiro Ogawa [Yamaguchi Medicine Vol. 9, pp. 571 to 576 (1960)] was injected intravenously with mature rabbits by dissolving extracts of physiological saline, terrestrial ethyl alcohol, or acetone extracts in physiological saline. The blood pressure drop was recognized.

⑦The "Chinese Medicine Dictionary", Kwon So-Shin Medical Institute, 1980, published by Shanghai Science and Technology Publishing Co., Ltd., page 2112. , Cat or chronic deity. When administered to mice with blood pressure, a moderate and sustained blood pressure drop was observed. Intravenous injection of the dragon dragon concentrate into dogs or cats under anesthesia rapidly reduced blood pressure. However, it has been reported to have no effect in oral administration or clinical application. In addition, on page 2114 of the back magazine, 10 ml of earth dragon periodicity (immersed 409 in 60 ml of ethyl alcohol at 60 ° C.) of 40% in concentration was applied every 10 ml, three times a day (that is, equivalent to 12 g / day of earth dragons). Take it. If you can not eat regularly, take the pure Jiryong powder in water to make a pill (add a small amount of excipient) and take 3 to 4g each time, 3 times a day (equivalent to 9 to 12g / day of earth dragons). Continued to take 30 to 60 days is effective for primary hypertension. In addition, ₂ ml each time the terrestrial B1 liquid (HgCl ₂ was used to remove hypoxanthin and the ion-exchange resin was used to separate the pressure-lowering component) (containing 8 g of the Chinese herb dragon), three times a day. Equivalent to 24 g / day (converted by the inventors)] It is reported that it was effective for essential hypertension.

Conventional customs of the production of earthworms or dry powder is roughly as follows.

(i) A method of dividing the earthworm's abdomen, removing the contents of the intestine (intestines and nito) and drying them in the sun, shade or low temperature (typically below 50 ° C).

(ii) A method of killing earthworms in vegetation and then removing the ashes and drying them in the sun, shade, or low temperature (typically 50 ° C or less) to obtain what remains intact in the earthworm body.

(iii) The earthworm is a method of removing nito from the body and drying it in vegetation or fire.

These dried products were used after being pulverized when necessary or used. Their method is simple and economical and has the advantage of being easy to implement at home. However, the dried or dried powder of the earthworms obtained by these preparations is within about 6 months when stored in a refrigerator at 0 to 5 ° C. or at room temperature at 5 to 45 ° C., and within a year when stored in a closed state. There is a drawback in which mold develops and becomes unusable.

Earthworms dried with nito full in the body as in the method of (ii) above, or earthworms dried in vegetation or fired ash as in the method of (iii), are extracted from hot water in most cases when used as a medicine. Or decoction with boiling water is often filtered to take the filtrate. In particular, the dried or dried powder of the earthworm by the manufacturing method of (ii) is rarely taken as a regular or powdered earthworm, or as a pill or the like.

The earthworms according to the method of (i) are often taken as hot water immersion liquids or liquid solutions, in addition to regular or earthworm powders, or by adding a small amount of water or a small amount of excipients to the powder. According to the manufacturing method of (ii) and (iii), the yield of the dry earthworm of 10 to 16% of moisture from the mountain earthworm is 5 to 9%, and the yield of the manufacturing method of (ii) is 13 to 19%.

Recently, one of the inventors of the present invention has reported a health food or a manufacturing method thereof [Japanese Patent Application Laid-Open No. 59-216572 (Dec. 6, 1984)]. This manufacture is a great way to get earthworm dry powder as a health food. However, it has been found to be insufficient in terms of its efficacy as a method of manufacturing earthworm dry powder for the purpose of medicinal use such as atherosclerosis (nickname, antihyperlipidemic), diabetes treatment and blood pressure control. In other words, only external soil cannot be selectively removed when an external action is performed to remove the excrement remaining in the living body of the earthworm. No matter how careful the operation, it was found that the visceral and bodily fluids containing a lot of ingredients that play an important role are removed along with the soil, resulting in insufficient efficacy. In addition, the yield for mountain earthworms is less than 10 to 19%. In addition, the major problem is that since the vacuum drying of the final finishing process is operated for a long time over 80 hours at 80 ℃, 0.3 torr vacuum, it was found that the destruction or activity of enzymes in the earthworm dry powder, which plays an important role in pharmacology and drug efficacy. Therefore, compared with the earthworm dry powder obtained by the manufacturing method of this invention, it turned out that the drug efficacy of the hypertensive agent of the earthworm dry powder obtained by Unexamined-Japanese-Patent No. 59-216572 declines rapidly. Vacuum drying of the final finishing process was performed under the same conditions as those described in the publication. In other words, the blood pressure lowering effect of the earthworm dry powder of the product obtained by varying the heating time to 20, 24, 26 and 30 hours with a vacuum degree of 0,3 Torr at a temperature of 80 ° C. was measured by the method of Pharmacological Test 1. Compared with the earthworm dry powder of the manufacturing method of this invention, it turned out that the drug efficacy is rapidly lost by about 48, 77, 82, and 90%, respectively.

Recently, one of the inventors of the present inventors, Hisashi Mihara and other co-researchers, the active material for earthworms has an appropriate pH of 8 to 10, a stable pH of 5 to 10, trajilol (trade name), transamine (trade name), and soy trypsin. It was confirmed that it is an enzyme protein that is inhibited by inhibitors and serum, and has a composition having a plasminogen activating action and a fibrin soluble action and a fibrinogen soluble action.

Japanese Patent Application Laid-open No. 58-148824 (Application Date 2, 1982, 1982) of the preparation method of the active enzyme substance consisting of crude enzyme protein fraction by extraction of aqueous solvent from earthworm and purified enzyme protein fraction by purification thereof. 27) and US Patent Application No. 470394 (filed Feb. 28, 1983), and US Patent Application No. 8305359 (filed Feb. 25, 1983) for a foreign application based on its priority claims. 47795 (filed February 25, 1983), French Patent Application No. 03165 (filed February 25, 1983), Federal Republic of Germany Patent Application No. P3306944.1 (filed February 28, 1983) and Canadian patent application 422034 (filed February 21, 1983) is reported.

Mihara Hisashis, filed a patent application for six novel proteases from earthworms, ie Japanese Patent Application Publication No. 59-63184 (filed date Digestion 57 (1982) October 2) and their proteases. Japanese patent application publication No. 59-184131 (filed date March 58, 1983) and a patent application for a current dissolving agent as an active ingredient and a foreign patent application that claims priority of their combined application, that is, a Korean patent The following patent applications are reported, including application 2900 (filed June 30, 1983) [AU.P.App.16293 (filed June 27, 1983), CA.P.App. 431387 (filed June 28, 1983), DK.P.App.3008 (filed June 29, 1983), EP.P.App, 83106288.0 (filed June 28, 1983), ES.P.App 523754 (filed June 30, 1983), FI, P.App.832383 (filed June 29, 1983), NO.P.App.2399 (filed June 30, 1983), PH.P. App. 29151 (filed June 30, 1983), TW.P.App. 7211983 (filed June 18, 1983), US.P. App. 508163 (filed June 27, 1983).

In the investigation results of the present inventors, there was no document which reported the action activity of the antihypertensive agent which contains the guin, earthworm, and earthworm dry powder. Moreover, when the earthworm dry powder was administered to a patient with high blood pressure and / or low blood pressure, there was no document reported on the blood pressure regulating action of the earthworm dry powder such as bringing the highest blood pressure and the lowest blood pressure to the normal level.

A blood pressure lowering agent and a blood pressure raising agent are considered to act separately. In other words, the blood pressure lowering agent has a function of lowering blood pressure and the blood pressure raising agent has a function of raising blood pressure, so it should always be taken under the supervision of a specialist. For example, in the case of a blood pressure lowering agent, when a certain hypotensive effect is confirmed, treatment is performed such as changing the type of drug or temporarily stopping taking the drug according to the stage. When blood pressure rises again, you must resume taking the medicine substitute for it. It is desirable to be able to treat patients with hypertension and / or hypotension with one agent. In other words, there is a need for a medicament that brings the highest and lowest blood pressure to high blood pressure and / or low blood pressure patients. In addition, the antihypertensive blood pressure agent is accompanied by a long-term dose, so a safe and free of side effects is needed. For this reason, the present inventors have been intensively researching for many years for the purpose of creating a high-safety medicine with no side effects from natural medicines.

It is known that the building of the earthworm has a blood pressure lowering action, as indicated in the above prior document. However, the yield of the earthworm dried by the conventional conventional technique is not only 5 to 19% less than the mountain earthworm, but also mold is generated or deteriorated within one year even when stored in a sealed state at room temperature of 5 to 45 ℃ as medicinal Becomes inadequate. Insufficient or insufficient efficacy or side effects of the drug, such as the destruction or loss of the enzyme in the earthworm dry matter. Therefore, in the present invention, no destruction or activity of enzymes in the earthworms is dried and no side effects such as destruction of erythrocytes, that is, hemolysis and tachycardia, are high in safety, and can be stored or stored for at least 4 years in a closed state. The dry powder of the earthworm was studied in detail to obtain a high yield.

As a result of detailed studies to solve the above problems, a novel and progressive improvement method of the earthworm dry powder shown below was identified.

[Preparation 1:]

1 part (weight) of mountain worms at a temperature of 1 to 30 ° C. for 10 to 30 hours, preferably 10 to 30 hours at a temperature of 4 to 28 ° C., in 1 to 10 parts of fresh water (capacity), or acetic acid, citric acid, succinic acid, 0.3 (%) or less, preferably 0.1 (%) or less, containing at least one compound of an organic acid such as malic acid, tartaric acid, lactic acid, or an inorganic acid such as phosphoric acid, sulfuric acid, hydrochloric acid, or sodium or potassium salt of these acids 1 to 10 parts (capacity) of an acidic aqueous solution of low concentration or pH 3 to 6.5, at a temperature of 1 to 25 ° C for 0.5 to 72 hours, preferably at a temperature of 2 to 15 ° C for 1 to 40 hours After excretion, the soil in the digestive tract of the mountain earthworm was sufficiently excreted, and then the soil attached to the surface of the body of the earthworm was washed with water, and then simply drained and crushed the mountain earthworm as it was.

The thick suspension of the obtained earthworms was frozen at a low temperature of -5 ° C or lower, preferably -10 to -60 ° C, and then freeze dried and vacuum dried. The conditions of freeze-drying and vacuum drying are 5-100 hours, preferably 10, while gradually raising the temperature with a vacuum of temperature -60 to + 90 ° C and a vacuum degree of 100 mmHg or less, preferably -40 to + 80 ° C or less and 30 mmHg or less. Freeze-drying and vacuum drying were performed for 60 hours to obtain dry powder of aseptic earthworm.

[Manufacturing 2:]

1 part (weight) of the mountain worm after washing off the dirt attached to the body surface with water is 10 to 30 hours at a temperature of 1 to 30 ℃, preferably 10 to 30 hours at a temperature of 4 to 28 ℃, fresh water 1 to 10 ℃ 0.3 parts by weight or less of an organic acid such as acetic acid, citric acid, succinic acid, malic acid, tartaric acid, lactic acid or inorganic acids such as phosphoric acid, sulfuric acid, hydrochloric acid, or at least one compound of sodium or potassium salts of these acids 1 to 10 parts (capacity) of a low concentration or preferably an acidic aqueous solution of pH 3 to 6.5 containing 0.1 (weight)% or less, at a temperature of 1 to 25 ° C. for 0.5 to 72 hours, preferably at a temperature of 2 to 15 ° C. After leaving for 1 to 40 hours in the earthworm itself to excrete the soil in the digestive tract of the earthworm by the excretion power of the earthworm itself, and then with or without washing with water, the earthworms are drained and pulverized as the earthworm It was.

The thick suspension of the earthworms obtained was frozen at a low temperature of -5 DEG C or lower, preferably -10 to -60 DEG C, and then lyophilized and vacuum dried. The conditions of the lyophilization and vacuum drying are 5 to 100 hours while increasing the temperature stepwise with a vacuum of -60 to + 90 ° C and a vacuum degree of 100 mmHg or less, preferably -40 to + 80 ° C and 30 mmHg or less. Freeze-drying and vacuum drying were carried out for 10 to 60 hours to obtain dry powder of aseptic earthworms.

As a method for crushing the earthworm, that is, the method of destroying the tissue (cells) as it is, it is preferable to use a homogenizer, a blender, a homomixer, a lightning rod, and a device of a pressurized cell destruction device to prepare a suspension or homogenate. The temperature of this grinder is 1 to 25 ° C, preferably 2 to 15 ° C.

Yellow-brown or brown-earthworm dry powder was obtained in a yield of 20 to 35% from the mountain earthworm by the tissue. Typically, the earthworm dry powder has 5 to 16% moisture, preferably 7 to 14% ash, 3 to 8% ash, preferably 4 to 7%, nitrogen to 1 to 11%, preferably 6 To 11%. In the earthworm dry powder, 18 kinds of aspartic acid, threonine, serine, glutamic acid, florin, glycine, alanine, cystine, valine, ethionine, isoleucine, leucine, tyrosine, phenylalanine, tryptophan, lysine, histidine, arginine or It contains amino acids before and after.

[Test Example 1]

Results of the schematic analysis of the earthworm dry powder products (M-1 to M-9) obtained by the method of the above-mentioned Production method 1 (late Examples 13 to 16) and production method 2 (late Examples 17 to 21). Is shown in Table-1. Earthworm dry powder product symbol M-1 by the manufacturing method 1 was obtained by the method of Example 13. Similarly, M-2 was obtained in Example 14, M-3 in Example 15, and M-4 in Example 16. The earthworm dry powder product symbol M-5 by the manufacturing method 2 was obtained by the method of Example 17. Similarly, M-6 was obtained in Example 18, M-7 in Example 19, M-8 in Example 20, and M-9 in Example 21.

TABLE 1

[Test Example 2]

Table 2 shows the results of component analysis of M-2 and M-4 of the earthworm dry powder product obtained in the method of Preparation Method 1 and M-5 of the earthworm dry powder product obtained in the method of Preparation Method 2.

TABLE 2

[Test Example 3]

Amino acid analysis of crude protein in earthworm dry powder products of M-2 and M-4 obtained by the method of Preparation 1 and M-5 obtained by the method of Preparation 2, and comparison with the analysis of fish meal and soy flour of protein food One result is shown in Table-3.

TABLE 3

* Cited from data described in P. McDonald, R. A. Edwards and J. F. D. Greenhalgh: Animal Nutrition (1973).

The earthworm dry powders obtained by the methods 1 and 2 in Tables 2 and 3 were found to contain a sufficient amount of crude protein, crude oil, and various metals.In addition, the amino acid composition of the crude protein contained a large amount of essential amino acids. okay.

The inventors of the present invention are to remove dirts in the mud, straw, fallen leaves, clays and soils in the digestive tract as soon as possible, and to improve the yield, physicochemical activity, and preservation of earthworm dry powder products. It was found to be an important operation to raise the back.

In particular, excrete soil in the digestive tract of the earthworm first. The manufacturing method 1 which wash | cleans and removes the dirt which adheres to the body surface was able to complete a washing | cleaning operation in shorter time than the manufacturing method 2.

As a property of earthworms, when handling a large number of mountain earthworms, the earthworms are entangled with each other and collected. If you try to clean and remove the dirt attached to the surface of the body before discharging the digestive tract, the earthworm will excrete the soil in the digestive tract during the operation and attach it to the body surface of the earthworms. It doesn't work. In order to clean, it is necessary to repeat washing 4 to 5 times as shown in Example 17-21 mentioned later. As a result, there is a loss of the earthworm, the body fluid from the wound of the living body and the digestive fluid which are accompanied by the increase of the number of washing operations.

However, thereafter, when left in the above aqueous solution or fresh water to excrete the remaining soil, there is a relatively small amount of distribution attached to the body surface of the mountain earthworm. Therefore, when it is a small quantity, it may be good for light cleaning in the subsequent drying process, or it may be unnecessary.

Although the same type of red earthworms (Lumbricus rubellus) are used, and the cleaning method is different from the manufacturing method 1 and the manufacturing method 2, the yield of the earthworm dry powder product obtained by the operation of making the same conditions afterwards and the outline analysis result of the product ( A comparison of the above Table-1 and Table-2) is shown in Table-3-1.

Table 3-1

In other words, as can be seen from the comparison of the products M-3 and M-5, the comparison of the products M-4 and M-6 and the comparison of the products M-2 and M-7 shown in Table-3-1, The earthworm dry powder product obtained in the above-mentioned products had a yield of 0.5 to 1%, a noble content of 0.7 to 0.9%, and a nitrogen of 0.4 to 1.4% than the product of Preparation 2. In addition, the product of the manufacturing method 1 obtained the preferable product which tends to have a slightly higher physiological-chemical activity (refer to the result of Table-5 of the specification) than the product of the manufacturing method 2, and has few side effects. In addition, it is better to leave the mountain worms in an aqueous solution containing at least one compound of the above-mentioned organic acid or inorganic acid or the sodium or potassium salt of the mass production thereof in a low concentration as described above. Excretion is fast and excretion rate is large.

In the said manufacturing method 1 and the manufacturing method 2, the completion partner of the earthworm dried product after completion | finish of drying is often in the state of aggregation or agglomeration. However, when this is put into a grinder, it becomes powder easily. When administering the earthworm dry matter to mammals, including humans, it is preferable to powder than lumps.

Mihara Hisashis (Japanese Patent Application Laid-Open Nos. 59-63184 and 59-184131) fractionated six new proteases of a gene for enzyme from earthworms. That is, Mihara Hisashi was subjected to yuan fractionation of the supernatant, which had been incubated for 2 days by adding 10 times the amount of physiological saline to the earthworm dry powder, and the precipitate was subjected to gel filtration by Sephacryl S 200, and the obtained protein. DEAE-cellulose-ion exchange chromatography was performed on the fractions to obtain proteins of I, II, III and fractions having casein degradation and fibrin degradation activity. The fractions of I, II, and III were further purified by DEAE-cellulose, Sephadex G-75, Toyota WH 55, ACH-Sepharose, Benzamidine-Sepharose and the like to obtain 6 fractions of purified enzyme. Molecular weight was determined by SDS-PAGE, and the molecular weight of fraction 1-0 was the lowest, calculated as 23,500, and subsequently the molecular weight was increased to 1-1,1-2, II, III-1, III-2 and III-2. Has a molecular weight of 34,200. The six-point coin point was measured by isoelectric point electrophoresis, with 1-0 being the highest and pH 4.12, followed by a lower pH and a pH of 3.52 at III-2.

These six fractions are new proteases that are also different from serine enzymes, and their six fractions have a suitable pH of around 8 or pH 8-10, a stable pH of 4-12 or 5-12, and a suitable temperature of It is reported that 50 ° C. or 50 to 60 ° C., and loss of activity conditions were 70 minutes at 70 ° C.

10 times the amount of physiological saline was added to the dry powder products M-4 and M-5 of the earthworms obtained in Preparation Methods 1 and 2 above, and the supernatant thereof was measured on a standard fibrin plate, and as shown in Table 4, the immediate graft activity was decreased. Admitted. The physiological saline solution of the earthworm dry powder of M-4 was incubated at 37 ° C. As shown in Table 4, the supernatant activity of the supernatant was about 4 times at 10 days and 5 times at 5 days and 75 days. The fact is that the protease of this protease is present in the freeze-dried powder of earthworms, and it is thought that there is an expression of enzymatic activity by self-extinguishing.

The activity of the supernatant on the 50th day was calculated to be about 8,000 IU / ml in terms of international units of urokinase. This enzyme is also plasminogen. Plasminogen on the side of the standard fibrin plate by dissolving with the free fibrin plate and the standard fibrin plate. Glass. Prasminogen with enzymatic activity that directly dissolves fibrin and has a large lysate compared to the plain tube. Activator activity was also shown.

In the above operation, the result of using the earthworm dry powder of M-5 instead of M-4 was the same as in Table-4.

Table 4

10-fold physiological saline was added to the earthworm dry powder of the present invention and the supernatant activity of the supernatant when incubated at 37 ° C.

The reason why the present invention (i.e., earthworm dry powder, meaning the following) orally administered to rats and humans shows blood pressure control effect, that is, antihypertensive and / or antihypertensive effect, is unknown or earthworm dry. It is thought to be due to the action of the protease (protein) itself or enzymes thereof contained in the powder or precursors (proteins) of these enzymes themselves or proteins or lipids or their unknown compounds themselves or their combination. The nitrogen content of the present invention most preferred as a blood pressure regulator or antihypertensive agent is 7 to 10%, that is, crude protein content of 43.8 to 62.5%.

Next, a preferred specific example of the freeze-drying and vacuum-drying operation of the suspension by crushing the earthworm obtained in the above-described production methods 1 and 2 is as follows.

The ground mass of the earthworm, ie the suspension of the earthworm, is frozen at -10 to -60 ° C, preferably at -30 to -50 ° C for 5 to 60 hours, and then at the same temperature under vacuum of 0.01 to 0.2 mmHg for 5 to 12 hours. Freeze-drying. Next, it is dried under vacuum at 0.01 to 0.2 mmHg for 5 to 15 hours at 20 to 30 ° C.

Next, it is dried for 10 to 20 hours under a vacuum of 35 to 50 DEG C and 0.1 to 0.5 mmHg. The vacuum drying of the final finishing process is sterile when vacuum dried at 0.01 to 0.5 mmHg under vacuum at 70 to 80 ° C., preferably at 75 to 80 ° C. for 5 to 10 hours, and contains 5 to 15% moisture. You can get In particular, vacuum drying of the final finish is an important operation. In order to finish the dry powder of the earthworm in a sterile state without losing the activity of the protease and the precursor of the enzyme contained in the earthworm dry powder, the present inventors studied in detail lyophilization. The operating conditions described above were found to be the important condition that the combination of the three factors of the vacuum degree (0.01 to 0.5 mmHg), the heating temperature (70 to 80 ° C.), and the time (5 to 10 hours) of the vacuum drying condition of the final process of vacuum drying was important. Was established.

As indicated above, all six kinds of purified protease from the earthworm dry powder (Japanese Patent Application Publication Nos. 59-63184 and 59-184131) have been reported to lose their activity at 70 ° C. for 60 minutes. , Protease in the earthworm dry powder obtained in the production method of the present invention does not lose activity as shown in Table-4. This reason is as follows.

In the method of the above-described Japanese Patent Application Laid-Open Publication No. 6, the proteolytic enzymes were deactivated by heating at 70 캜 for 60 minutes under atmospheric pressure (1 atm). However, in the final process of the manufacturing method of the present invention, due to heating at 70 to 80 ° C. for 5 to 10 hours under a high vacuum of 0.01 to 0.5 mmHg, proteolytic enzymes in the earthworm dry powder did not lose their activity. In other words, as a preferable condition for vacuum drying of the final finishing process of the earthworm dry powder of the present invention, the combination of the heating temperature, the heating time and the vacuum degree becomes an important condition for sterilizing various bacteria without losing activity of the protease. These combination conditions are conditions established by the present inventors and the like.

In the case where the earthworm dry powders obtained in the methods of Preparation Methods 1 and 2 were stored at room temperature of 5 to 45 ° C. for 5 years in a sealed state, mold occurrence, other physical properties, and chemical alterations were not recognized at all.

The earthworms used in the present invention include red earthworms (Lumbricas rubellus), LT earthworms (Lumbricuster-restris), earthworms (Eiseniafoetida), brown fishing earthworms (Allolophora caliginosa), purple fishing earthworms (Dendrobaena octaedra), Earthworm (Allolophora japonica Michael-sen), Hotda earthworm (Drawida hattamimizu Hatai), etc. Horst), earthworm worm (Pheretima hilgedorfi), worm earthworm (Pontodrilus matsus-himensis Iizuka), earthworm (Tubifex hattaiNomura), wall paulownia earthworm (nickname worm earthworm) [Limnodrilus gotoi Hatai = L. Also, any earthworm can be used if it is not poisonous.

The earthworm dry powder of the present invention may be in an oral or parenteral form when administered for clinical treatment, but is preferably orally administered. As an oral form of the present invention, a capsule, tablet, granule, powder (powder), coating agent, dragee, emulsion or the like is used by mixing with the present invention or an appropriate pharmacologically acceptable pharmaceutical carrier.

Examples of the pharmaceutical carrier include, for example, lactose, white sugar, mannitol, glucose, starch, sorbitol, glycine, calcium phosphate and microcrystalline cellulose; Starch, gelatin, arribia rubber, glucose, white sugar, sorbitol, mannitol, tragant, hydroxypropyl cellulose, hydroxypropoxymethyl cellulose, carboxymethyl cellulose, 2-methyl-5-vinylpyridine-methacrylic acid- Methyl acrylate ethyl copolymer, polyvinyl pyrrolidone, sodium arginate and the like; Examples of the lubricant include stearic acid, hydrogenated oil, magnesium stearate, calcium stearate, polyoxyethylene monostearate, talc, silicon oxide, polyethylene glycol, and the like; Starches containing potato starch, surfactants, etc. as disintegrating agents; Examples of the humectant include sodium lauryl sulfate. In addition, when administered parenterally, it can be used as a suppository. In particular, cacao butter, witemsol, subanal, polyethylene glycol, polypropylene glycol, glycerogelatin, gelatin capsules, etc. are used as the base of the suppository. In addition, well-known safe preservatives, such as methyl para hydroxy benzoate, propyl para hydroxy benzoate, butyl para hydroxy benzoate, and butyl hydroxy anisole, are used in combination.

The dosage of the earthworm dry powder of the blood pressure control agent or the antihypertensive agent of the present invention may vary depending on the administration method, the age, weight, condition, and type of disease of the patient. . Most preferably, the dosage is divided into 1 to 3 times a day from 0.02 g to 2 g per day.

[Action:]

The toxicology and hypertension treatment effect of the earthworm dry powder of the present invention or the pharmacological test method of blood pressure drop and the results thereof will be described in detail below.

[A. Acute Toxicity Test:]

Acute toxicity test by oral administration was performed using 5 groups of 30 mice each weighing 30 ± 2 g and male Wistar males weighing 100 ± 2 g each. Earthworm dry powder M-1 of the present invention (water 10.2%, ash, 5.1%, nitrogen containing 9.4%); Copper M-2 (containing 10.4% moisture, 5.3% ash, 8.6% nitrogen); Copper M-3 (containing 10.7% moisture, 5.2% ash, 9.2% nitrogen); Copper M-4 (containing 10.6% moisture, 5.6% ash, 9.6% nitrogen); Copper M-5 (containing 9.5% moisture, 4.5% ash, 7.8% nitrogen); Each dose of copper M-6 (containing 9.2% moisture, 4.7% ash, 8.4% nitrogen) was increased from 0.1 g / kg to 8 g / kg to the mice (0.1 to 5 g / kg) and rats (2 to 8 g / kg). kg) were individually administered by forced administration with a throat tracheal rod. Animals were maintained at an animal room temperature of 22-23 ° C. during the test period and observed for 14 days after dosing. No deaths from the doses administered were recognized at all. Post-dose intoxication and behavior were observed over time, but no difference was observed between normal animals and hadong.

In addition, weight gain was almost no difference from normal animals. No macroscopic disturbances were recognized in any part of the major organs in the post-test autopsy. Therefore, the earthworm dry powder of the present invention could not determine the LD50 value because of its very low toxicity.

B. Actions on naturally occurring hypertensive rats (hereinafter referred to as SHR):

[Pharmacology Test 1]

Blood pressure drop and heart rate test for SHR:

100 mg / kg (M-4) of the earthworm dry powder (M-4) of the sample suspended in 0.5% of carboxymethyl cellulose solution to male SHR of 10-12 weeks old (200-300 g in weight, 150-200 mmHg in blood pressure), which was choked one night. Hydralazine hydrochloride (10 mg / kg) was prepared orally, and blood pressure was measured at 0, 1, 2, 4, and 6 hours (average of 6 to 7). That is, the blood pressure lowering action and the heart rate increase were determined by comparing the blood pressure and heart rate before administration of the specimen with the blood pressure and heart rate after administration of the specimen.

The blood pressure was measured by heating the SHR at 45 to 50 ° C. for about 5 minutes in advance, and then using the automatic blood pressure recorder, using the tail plethysmograph method [the journal, the laboratory, and the cliché. Nikal. Magazine, vol. 78, p. 957 (1971), measured non-invasively the peak blood pressure (constrictor blood pressure) of the tail artery. The test results are shown in Table-5.

The highest blood pressure difference before and after administration of the sample was expressed as a blood pressure lowering effect. By administering 100 mg / kg orally of the earthworm dry powder of all the samples of the product obtained in the method 1 (M-2 and M-4) and the product obtained in the method 2 (M-5 and M-6) to SHR. , Maximum coercive action was observed at 1 to 2 hours after administration. That is, the blood pressure of this SHR was lowered by 28 to 35 mHg and the action was continued for 6 hours.

In addition, the results of measuring the increase in heart rate are shown in Table-5. It was found that the dry powder of the earthworms of all specimens did not have undesirable side effects on the heart, such as tachycardia, like hydralazine hydrochloride.

TABLE 5

a) Each value represents the mean ± standard deviation (Mean ± S.D) of 6-7 SHR.

b) Significant difference from the value of the control group only in 0.5% aqueous solution of carboxymethyl cellulose: ^* P <0.05, ^** P <0.01, ^*** P <0.001

[C. Hemolytic action]

[Pharmacological examination 2]

5 parts of physiological saline to 1 part (weight) of the dry powder of the earthworm of each of the four kinds of products (M-2 and M-4) obtained in Production Method 1 and the products (M-5 and M-6) obtained in Production Method 2 (M-5 and M-6). After the suspension was added well, the mixture was allowed to stand for 24 hours in a refrigerator at 5 ° C., filtered under reduced pressure, and the filtrate was used as a water extract.

Ethyl alcohol or acetone extract component was added to 1 part (weight) of dry powder of four kinds of specimens by adding 5 parts (weight) of etaal alcohol or acetone, and then suspended well, and then left at 15 to 20 ° C for 24 hours. Filtration under reduced pressure and the filtrate was dried by drying under reduced pressure at 40 to 45 ° C. A 30% solution was prepared with physiological saline to which 1% carboxyethyl cellulose was added. The hemolytic action test was performed by the following method. Blood was collected from the ear vein of a mature rabbit having a weight of about 3 kg and subjected to de-fiberization to obtain de-fiberized blood. Dilute each portion of the water extract, ethyl alcohol and acetone extract to 5, 1, 0.1, 0.01, 0.001, 0.0001% by weight with physiological saline, and place them in a 5 ml small test tube and pipet the defiberized blood by pipette. 1 hour after the dropwise addition, hemolysis results were determined. As shown in Table 6, in all cases, hemolysis was not recognized in all samples of the earthworm dry powder.

TABLE 6

Prior art documents report that the earthworm poison has a destruction of red blood cells, that is, hemolysis in the earthworm building (earthworm). [Earth and life: Ohfuchi Shin Ryuuser, October 30, 1947, published by Makki West, 223 Pages 226, and reprinted earthworms: Hatay Shinjigeru, April 30, 1980, published by Scient Co., Ltd., pages 160 to 163, and the water extract and ethyl alcohol extract of the earthworm dry matter (Jelong) Although it has been reported to have a hemolytic action (Kenji Igawa, Vol. 9, Yamaguchi Medical Vol. 9, pp. 571 to 576 (1960)), it has been found that the earthworm dry powder according to the production method of the present invention does not have such a distinctive action.

[D. Oral Administration of Antihypertensive Drugs in Humans of Earthworm Dry Powder:]

Applicants to 14 volunteers (7 males and 7 females) of 8 hypertension patients (4 men and 4 women) and 6 hypotension patients (3 men and 3 women) Capsule A [1 capsule prepared in Example 4 containing earthworm dry powder (M-2 moisture 10.4%, ash 5.3%, nitrogen 8.6%) 150mg, capsule E [1 capsule prepared in Example 11 Earthworm dry powder (M-4, water 10.6%, ash 5.6%, nitrogen 9.6%) 150 mg and capsule F prepared in Example 12 [1 capsule is an earthworm dry powder (M-5, Water 9.5%, ash 4.5%, nitrogen 7.8%) 150mg] once capsules once, three times a day orally was taken within 30 minutes after breakfast, lunch, and dinner. After the start of administration, oral administration was carried out for 2 to 11 months for 2 months and then once every month for 1 month. The results showed that when administered to patients with hypertension as shown in Table-7, the highest and lowest blood pressures fell approximately equal to 30 mmHg. That is, the highest blood pressure of the hypertension patients was 30 to 39mmHg, the lowest blood pressure was 26 to 34mmHg. After the blood pressure reached a normal level, normal maintenance was recognized for a period of 3 to 7 months. In addition, when administered to patients with hypotension, the blood pressure was increased by 15 to 30 mmHg. That is, the highest blood pressure of the hypotensive patient was increased by 20 to 30 mmHg, and the lowest blood pressure was increased by 15 to 22 mmHg. After reaching a normal level, the maintenance of blood pressure was recognized for a period of 3 to 5 months. In addition, the long-term administration of the earthworm capsule of the earthworm dry powder of the present invention showed no abnormal drop in blood pressure and / or abnormal rise in human blood, and no other side effects were recognized.

A blood pressure regulator or antihypertensive agent comprising the earthworm dry powder of the present invention as an active ingredient alleviates blood pressure lowering and / or blood pressure synergism in humans, manifests a moderate drop and / or elevated pressure, and treats hypertension. Except for these, it has proved to be a safe and prolonged drug as their prophylactic agent.

Table 7

Example 1

Tablet B

Tablets C

According to the above formulation, tablets of various weights were prepared by tableting the powder mixed well.

Example 2

Granule A

The dry powder of the earthworm, lactose, microcrystalline cellulose and corn starch were mixed well using the fluidized bed granulator according to the above formulation, and sprayed with a 5% aqueous solution of hydroxypropyl cellulose as a binder to obtain granules after low temperature drying.

Example 3

Granule B

Granule C

Granules were prepared by extractors of well mixed powders according to the above formulation.

Example 4

Capsule A

In the above formulation, components other than hydroxypropyl cellulose were mixed well using a fluidized bed granulation apparatus, sprayed using a 5% aqueous solution of hydroxypropyl cellulose as a binder, dried at low temperature, and then granulated. The granules were filled in 250mg of hard capsules to prepare hard capsules.

Example 5

Capsule B

The hard capsules were prepared by filling 250 mg of the granules C prepared in Example 3 in light capsules.

Example 6

Capsule C

The prescription was mixed well, and the mixed powder was filled into the gelatin capsule of No. 1 by 250 mg to prepare a capsule.

Example 7

Enteric tablets

The tablets were uniformly mixed according to the above formulation, and then prepared by a tablet press, and then coated with a coating agent of the enteric coating film shown below to prepare an enteric tablet.

Coating agent

Example 8

Presenter A

Powder B

Each of the above ingredients was mixed well and uniformly in a cone mixer to prepare a powder.

Example 9

Suppository A

Suppository B

A suppository was prepared by injecting a melt of each of the above-mentioned prescriptions into an aluminum mold and cooling the mixture.

Example 10

Capsule D

Mix well as prescribed above. This mixed powder was filled into 250 mg of gelatin capsules of No. 1 to prepare a capsule.

Example 11

Capsule E

The prescription was mixed well and the mixed powder was laminated to the gelatin capsule of No. 1 by 250 mg to prepare a capsule.

Example 12

Capsule F

Mix well as prescribed above. This mixed powder was filled in 250 mg each of gelatin capsules of No. 1 to prepare a capsule.

Example 13

1 kg (approximately 2,000 earthworms) of lightly cleaned earthworms (red earthworms) are left for 4 hours at 18 ° C in an acidic aqueous solution of pH 6.2 in which a 1: 1 mixed acid of malic acid and citric acid is dissolved. After excretion, the earthworms were washed well with water to wash away the dirt and dust attached to the body surface of the earthworms.

Next, it was put into a mixer and wet-pulverized. The obtained suspension of the earthworms was put in a tray and frozen at -30 ° C for 40 hours, and then freeze-dried at -40 ° C for 6 hours under a vacuum of 0.1mmHg. Then, the temperature of the tray containing the tray was raised to 30 ° C and a vacuum of 0.1mmHg was obtained. After vacuum drying for 6 hours, the temperature of the shelf was then raised to 50 DEG C, followed by vacuum drying for 10 hours under a vacuum of 0.2 mmHg, and then the temperature of the shelf was raised to 80 DEG C, and dried for 8 hours under a vacuum of 0.2 mmHg. 280g of dry powder products (M-1) of an earthworm were obtained.

Example 14

1 kg of a light earthworm (red earthworm) washed with water was left in 3 l of an acidic aqueous solution of pH 5.5 containing phosphoric acid, tartaric acid, and lactic acid 1: 1 mixed solution at a temperature of 10 ℃ for 2.5 hours to excrete the soil in the digestive tract sufficiently. Afterwards, the earthworms were washed well with water to wash away the dirt and dust attached to the body surface of the earthworms.

Then it was wet milled with a mixer. The suspension of the earthworms obtained was placed in a tray and frozen at -25 ° C for 20 hours, and then the product temperature was lowered to -35 ° C, freeze-dried under vacuum of 01 mmHg for 7 hours, and then the temperature of the shelf on which the tray was loaded was 28 ° C. The worm-dried powder product (M-2) by vacuum drying for 10 hours under vacuum of 0.1 mmHg, followed by vacuum drying for 13 hours under vacuum of 0.2 mmHg at 40 ° C, and then drying for 8 hours under vacuum of 0.1 mmHg at 78 ° C. 275 g).

Example 15

1 kg of mountain worms (red worms) lightly washed with water were left in 3 l of fresh water for 16 hours at a temperature of 10 ° C. to excrete the soil in the digestive tract. The mountain earthworms were washed well with water to wash away the dirt and dust attached to the body surface of the mountain earthworms. Then, it was wet milled with an ultra homomixer. The suspension of the earthworms obtained was put in a tray and frozen at -25 ° C for 15 hours. The product temperature was then lowered to −35 ° C. and lyophilized for 6 hours under vacuum of 0.1 mmHg, followed by drying for 10 hours under vacuum of 0.08 mmHg at 30 ° C., followed by drying for 15 hours under vacuum of 0.2 mmHg at 40 ° C. Next, 245 g of earthworm dried powder product (M-3) was obtained by drying at 78 degreeC under the vacuum of 0.1 mmHg for 8 hours.

Example 16

1 kg of a mountain worm (red worm) lightly washed with water was left for 3 hours at a temperature of 13 ° C. in 2 L of an acidic aqueous solution of pH 5.8 in which malic acid was dissolved, and the soil in the digestive tract was thoroughly excreted. The mountain earthworms were washed well with water to wash away the dirt and dust attached to the body surface of the mountain earthworms. Next, it was wet-pulverized by the ultra homo mixer (made by Nippon Seiki Co., Ltd.). The obtained suspension of the earthworms was put in a tray and frozen at -30 ° C for 30 hours, and then freeze-dried for 8 hours under vacuum of 0.1 mmHg at a temperature of -30 ° C, and then the temperature of the shelf on which the tray was loaded was raised to 25 ° C, Vacuum drying at 0.1 mmHg vacuum for 7 hours, then raising the temperature of the shelf to 45 ° C., freeze drying for 8 hours under vacuum at 0.1 mmHg, and then raising the temperature of the tray containing the tray to 25 ° C. and vacuuming at 0.1 mmHg. Vacuum drying under 7 hours, then the shelf temperature is raised to 45 ° C. and vacuum dried for 12 hours under 0.1 mmHg vacuum, and then the shelf temperature is raised to 80 ° C. for 7 hours under vacuum of 0.1 mmHg 275 g of (M-4) was obtained.

Example 17

Dirt, straw, etc. attached to the surface of the body of 1 kg of earthworm (red earthworm) was washed well with water five times and washed. Next, the earthworms were left for 18 hours at a temperature of 15 ° C. in 2.5 L of fresh water to excrete the soil in the digestive tract. Next, the mountain worms were washed lightly, and then wet crushed with an ultra homomixer. The obtained suspension of the earthworms was put in a tray and frozen at -40 ° C for 24 hours. Next, the product temperature was lyophilized for 5 hours under vacuum of 0.1 mmHg at -40 ° C, then for 8 hours under vacuum of 0.1mmHg at 25 ° C, and then for 7 hours under vacuum of 0.1mmHg at 45 ° C. 240 g of dry powder product (M-5) were obtained.

Example 18

Dirt, powder, etc. attached to the surface of 1 kg of mountain worms (red earthworms) were washed well with water four times. Next, the earthworm was left in 2.5 liters of an acidic aqueous solution of pH 5.7 in which a mixed acid of malic acid and lactic acid 1: 1 was dissolved at a temperature of 15 ° C. for 2.5 hours to excrete the soil in the digestive tract. Next, the earthworms were lightly washed and then wet crushed with a mixer. The suspension of the earthworms obtained was put in a tray and frozen at -35 ° C for 24 hours. Next, the product was freeze-dried for 7 hours under a vacuum of 0.1 mmHg at a temperature of −35 ° C., followed by raising the temperature of the shelf on which the tray was loaded to 22 ° C. and vacuum drying for 10 hours under a vacuum of 0.1 mmHg. The temperature was raised to 42 ° C. and dried for 15 hours under a vacuum of 0.2 mm Hg, and finally, the temperature of the shelf was raised to 78 ° C. and dried for 7 hours under a vacuum of 0.1 mm Hg to obtain 265 g of a dry powder product (M-6) of an earthworm.

Example 19

Earthworms (Red Earthworms) Clean the dirt of mud, powder, straw, etc. on the surface of 1kg body five times with water, and then the earthworms contain a mixture of phosphoric acid, tartaric acid, and lactic acid 1: l: 1. The soil in the digestive tract was excreted by standing for 2.5 hours at a temperature of 14 ° C. in 3 L of an acidic aqueous solution of pH 5.3.

Next, the mountain earthworm was lightly washed with water, and then wet crushed with a homogenizer. Thereafter, the obtained suspension of the earthworms was put in a tray and frozen at -35 ° C for 30 hours, and then lyophilized for 7 hours under vacuum of 0.1mmHg at a temperature of -35 ° C, and then the temperature of the shelf was raised to 25 ° C to 0.1mmHg. After vacuum drying for 8 hours under vacuum, the dry powder of the earthworm is dried by raising the temperature of the shelf to 45 ° C. for 10 hours under vacuum of 0.1 mmHg, and finally drying the temperature of the shelf to 80 ° C. for 7 hours under vacuum of 0.1 mmHg. 265 g of product (M-7) was obtained.

Example 20

Earthworms (fishing earthworms) 1 kg of soil, dirt, straw, etc. attached to the surface of the body was washed well with water five times and washed. Next, the earthworm was left in 3 l of an acidic aqueous solution of pH 5.7 in which succinic acid was dissolved (in which 1 g of sodium acetate and 0.5 g of sodium sulfate was dissolved) at 13 ° C. for 2.5 hours to excrete soil in the digestive tract.

Next, the mountain earthworm was lightly washed with water, and then wet crushed with a homogenizer. Thereafter, 275 g of the earthworm dry powder (M-8) was obtained by freezing and vacuum drying the suspension of the earthworm obtained in the same manner as described in Example 18.

Example 21

Dirt, powder, straw, etc. attached to the surface of 1 kg of earthworms (coarse earthworms) were washed well with water five times and washed. The earthworms were then left for 2 hours at 15 ° C. in an acidic aqueous solution of pH 5.9 (with 0.7 g of potassium citrate dissolved therein) dissolved in a mixed acid of citric acid and tartaric acid 1: 1 to excrete soil in the digestive tract. . The earthworms were then washed lightly with water and then wet crushed with a blender. Thereafter, 283 g of earthworm dry powder (M-9) was obtained by freezing and vacuum drying the obtained suspension of earthworm in the same manner as in Example 18.

As described above, the present invention relates to a blood pressure control agent or an antihypertensive blood pressure agent, which is made of an earthworm dry powder as an active ingredient. The earthworm dry powder of the present invention has no side effects such as hemolysis and tachycardia and is safe and has an excellent blood pressure lowering action and a blood pressure increasing action, and has good storage or storage properties.

Subsequently, the volunteers of eight hypertension patients and six hypotension patients of 14 patients received the earthworm dry powder capsule (150 mg content) of the present invention once daily, three times daily, after breakfast, lunch, and dinner, orally 2 to 11 Administration was for months. The preparation of the present invention had the effect of lowering the blood pressure of the hypertension patient and raising the blood pressure of the hypotension patient to bring the blood pressure of the person to the normal level. Furthermore, the blood pressure reaching the normal level by the formulation of the present invention was seen to maintain normal blood pressure for 3 to 7 months. This dosing period, the ideal drop in human blood pressure and / or the manifestation of an ideal rise are neglected, and no other side effects were recognized.

In other words, the formulation of the present invention was found to be safe and excellent anti-hypertensive agents or antihypertensive agents or hypotensive agents.

Claims (8)

Mountain earthworms are left in an aqueous solution containing 0.3 (% by weight) or less of at least one compound selected from the group consisting of organic acid, inorganic acid, organic acid sodium salt, sodium acid salt, organic acid potassium salt and potassium salt of inorganic acid, or temperature 4 to 28. Leave the soil in the digestive tract of mountain worms for 10 to 30 hours in fresh water at ℃, wash and remove dirt attached to the body surface of mountain worms before and / or after this step with water, then crush and freeze-dry. And a blood pressure regulator comprising an earthworm dry powder obtained by vacuum drying as an active ingredient. Mountain earthworms are left in an aqueous solution containing 0.3 (% by weight) or less of at least one compound selected from the group consisting of organic acids, inorganic acids, sodium organic acids, sodium salts of inorganic acids, potassium salts of organic acids and potassium salts of inorganic acids, or temperatures of 4 to 28 ° C. 10 to 30 hours in the fresh water of the earthworm to excrete the soil in the digestive tract of the earthworm, and then remove the worms attached to the body surface of the mountain earthworm by removing water with the earthworm, or on the body surface of the mountain earthworm After cleaning and removing the attached soil, the earthworms are left in the aqueous solution or the fresh water to excrete the soil in the digestive tract of the mountain earthworms, and then the soils attached to the body surface of the mountain earthworms are washed out and removed with water. The suspension obtained by grinding as it is is frozen at -10 to -60 ℃, and then the temperature is stepwise in the range of -60 to 80 ℃ Freeze-drying and vacuum drying at a vacuum of 10 mmHg or less for 10 to 100 hours, and among these, the earthworm drying, which is dried for 5 to 10 hours under vacuum of 0.01 to 0.5 mmHg at 70 to 80 ° C. A blood pressure regulator containing powder as an active ingredient. According to claim 2, the earthworms are left in the aqueous solution or in the fresh water to excrete the soil in the digestive tract of the mountain earthworms, and then, the dirts attached to the body surface of the mountain earthworms are removed by washing and removing the earthworms as they are. And a blood pressure regulator for carrying out the above step. The blood pressure regulator according to claim 2, wherein the mountain earthworm before being left in the aqueous solution or the fresh water is a mountain earthworm in which a part of the dirt is washed with water. According to claim 2, after the dirt attached to the body surface of the mountain earthworm washed and removed, the earthworm is left in the aqueous solution or in the fresh water to excrete the earthworm in the digestive tract of the mountain earthworm, and the earthworm taken out as it is, A blood pressure regulator for carrying out the above steps. According to claim 4, after the dirt attached to the body surface of the mountain earthworm washed and removed, the earthworm is left in the aqueous solution or the fresh water to excrete the soil in the digestive tract of the mountain earthworm, and then washed with water, A blood pressure regulator for pulverizing the taken-out earthworm as it is and performing the subsequent steps. The blood pressure regulator according to any one of claims 2 to 5, wherein the organic acid is acetic acid, citric acid, succinic acid, malic acid, tartaric acid or lactic acid. The blood pressure regulator according to any one of claims 2 to 5, wherein the inorganic acid is phosphoric acid, sulfuric acid or hydrochloric acid.