KR930002010B1 - Antidiabetics - Google Patents

Abstract

No content.

Description

Diabetes treatment

The present invention relates to a therapeutic agent for diabetes. More specifically, the present invention relates to the treatment of diabetes with dry powder of earthworms and extracts of earthworms or earthworms having numerous blood sugar (hereinafter referred to as blood sugar) lowering activity and safety. The present invention also relates to a treatment and preventive agent for diabetes, which comprises an earthworm dry powder and l or earthworm extract as an active ingredient by a novel and progressive improvement method.

Earthworms have been used as medicines since they were ancient, mainly called earthworms and thoraxes in the Oriental Empire. The pharmacological and medicinal effects of the earthworm reported in the conventional literature are shown below.

① Earthworm and Life Obuchi Shinryu Published on October 30, 1947 (22 years old), Published by Makishobo, pages 223 to 226, and `` Reprinted Earthworms '' Daigi Kisage, April 30, 1980, Scientist, Inc. On pages 160-163, earthworms have a pharmacological action other than an agent for contracting stones in the bladder and excretion into the body, an agent for jaundice, a delivery agent, a tonic, a mother drug, a pill and an antipyretic agent. Invading the lineage, the other reported the destruction of red blood cells, ie hemolytic action.

(2) The Pharmacopoeia of the People's Republic of China Pharmacopoeia Committee of the People's Republic of China, 1977 edition, Part 1, pages 197-198. By convention, there are two kinds of tortoise products. One, Lumbricus Kwangtungesis, is torn the abdomen, washing the intestines, the soil and the sand, and drying them in the sun, in the shade or at low temperatures. The other worm (Lumbricus Nativus) is embedded in vegetation, killed, and then removed from ashes to dry in the sun, in the shade or at low temperatures, with the earthworm full of nito. The efficacy and indications of these two pterosaurs are: fever, calming the wind, calming the gasping and breathing paths (to improve blood circulation), farewell or convulsions caused by farewells, paralysis of the limbs, paralysis, paralysis, and joints. Swelling and pain, poor urination, gastrointestinal panting, hypertension, 4. 5-9gℓ day use is reported.

③ `` Our Chinese Herbal Medicine Series 3, Tohyong, Squid Bones '' Science Research of China on October 30, 1978, page 7, Matsuura Pharmaceutical Co., Ltd., on page 7, reports that there is a coercive effect on the tortoise frying (Ethyl alcohol extract of the tortoise). have.

(4) 「Natural Drug Dictionary」 Ondaengpyeon, April 15, 61, Digestion, Published by Hirogawa Bookstore, page 215 reports that the dragon is being used as an antipyretic, analgesic, diuretic and antidote.

⑤ Tanakamamoru [Hokkaido Medical Journal, Vol. 24, pp. 18-24 (1949)] extracts earthworms (without removing Ito from dried fragments) with aquatic water and adds ethyl alcohol to the concentrate of the extract. It was reported that the addition of precipitated substance (Lumbrofebrin) in ring gel seed solution and intravenous injection of this solution into cats under anesthesia resulted in a sudden drop in blood pressure and the promotion of blood coagulation in proportion to shock.

⑥ Igawa Genjiro [Yamakuchi Medical Vol. 9, pp. 571 to 576 (1960)] is an intravenous injection of a solution of physiological saline extracts of thorax, lysine ethyl alcohol or acetone extract dried in physiological saline to mature rabbits. I saw a drop in blood pressure.

⑦ `` Chinese Medicine Dictionary '' Heo-Gang Kang So-Shin Hagwon, 1980, published by Shanghai Science and Technology Publishing House, page 2112. Administration to cats or mice with chronic neonatal hypertension showed a slow and sustained hypotension. Anesthesia was injected intravenously in dogs or cats under anesthesia, and the blood pressure-lowering effect appeared rapidly. However, it has been reported that it was not effective in the case of dosing or clinical application. Again, page 2114 of Winter Solstice is taken with 40% of thorax simmer (immersing 40 grams of dinosaurs in 100 ml of 60-degree ethyl alcohol), 101 ml each time, three times a day (that is, equivalent to 129 liters of thorax). . If you can not drink ink, add pure water powder to water to make a pill (add a small amount of excipient), and take 3 to 4 g each time, three times a day (equivalent to 9 to l2 gℓ of dinosaurs (converted by the inventors)). . Continue taking 30 to 60 days is effective for essential hypertension. In addition, _{2 ml of} thorax dl liquid (except hippoxanthine using HgCl ₂ , separated from blood pressure lowering component using ion exchange resin) and 2 ml (containing 8 g of medicinal herb) per day It is reported that taking three times (equivalent to 24g 1 day of the dinosaurs (converted by the inventors)) was effective for essential hypertension.

Conventional customs for preparing earthworms or dried powders are roughly as follows.

Ⓘ Tore the earthworm's abdomen to remove the contents of the body (intestines and nito) and to dry in the sun, shade or at low temperatures (usually below 50 ° C).

지렁이를 초목을 태운 재안에 넣어서 죽인 다음, 재를 제거하고 천일, 그늘 또는 저온(통상 50℃ 이하)에서 건조하고 지렁이 체내에 니토가 차있는채로의 것을 얻는 방법.

Earthworms are killed by placing them in vegetated ashes, removing ashes, drying them in the sun, shade or at low temperatures (typically below 50 ° C), and getting nito in the earthworm body.

지렁이 체내의 니토를 제거한 다음, 초목을 태운 재 또는 불재안에 넣어서 건조시키는 방법.

A method of removing earthworms from the earthworms and then drying them in burnt or firewood.

And so on. These dried products were ground and used when needed or when used. Their manufacturing method has the advantage of being easy to implement at home in a simple or economic way.

However, the dried or dried powder of the earthworm 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 1 year when stored in a sealed state. There is a drawback that mold occurs and becomes unusable within a short period of time.

의 제법처럼, 체내에 니토가 차있는채로 건조한 지렁이 또는

의 제법처럼 초목을 태운 재 혹은 불재안에서 건조한 지렁이는 약으로서 사용할때에는 대개의 경우 열수로 추출 또는 자불수에 다린다음 여과하여 여액을 복용하는 수가 많다. 특히

의 제법에 의한 지렁이의 건조물 또는 건조분말은 토룡찡크 또는 분말인채 혹은 환약등으로하여 복용하는 일은 드물다. ⓘ의 제법에 의한 지렁이는 열수침지액, 곰액으로서 복용하는 외에 토룡찡크 또는 토룡 분말인채 혹은 분말에 소량의 물 혹은 소량의 부형제를 가하여 환약으로하여 복용하는 수가 많다. ⓘ 및

의 제법에 의할때, 산지렁이로 부터 수분 10∼16%의 건조 지렁이의 수율은 5∼9%, 동일하게

의 제법에 의할때의 수율은 13∼19%이다.Electricity

Like the recipe for a dry earthworm

In the case of using the dried earthworm in the firewood or firewood burned by vegetation as in the preparation of the method, the filtrate is usually taken by extraction with hot water or by filtration. Especially

It is rare to take dried earthworm powder or dry powder of earthworm by the method of the method as earthen mushroom or powder or pills. The earthworms prepared by the method of ⓘ are often taken as hot water immersion liquids or bear liquids, and are often taken as pills by adding a small amount of water or a small amount of excipients to the tortoise jink or tortoise powder. Ⓘ and

According to the manufacturing method of the dry earthworm with a moisture of 10 to 16% from the mountain earthworm is 5 to 9%, the same

The yield is 13-19% by the formula.

Recently, one of the inventors of the present invention, Ishiii Ichiichi, has prepared a health food or a manufacturing method thereof based on protein and lipids of earthworms (Japanese Patent Application Laid-Open No. 59-216572 (published date (December 6, 1984)). Reported. This method is an excellent way to obtain earthworm dry powder as a health food. However, it has been found that the method of manufacturing earthworm dry powder for the purpose of medicinal use such as atherosclerosis (nickname antihyperlipidemic agent), diabetes treatment agent and blood pressure lowering agent is not sufficient in terms of drug efficacy. In other words, when the external action is performed to remove the excrement remaining in the living body of the earthworm, only the soil can not be selectively removed. 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 together with the soil and thus the drug is insufficient. In addition, the yield for mountain earthworms is low as 10-19%. In addition, the major problem is that since the vacuum drying of the final finishing process is operated for a long time over 20 hours at a vacuum of 80 ° C and 0.3 Torr, it was found that the enzymes in the earthworm dry powder, which plays an important role in pharmacology and drug efficacy, were destroyed or inactivated.

Therefore, compared with the earthworm dry powder obtained by the manufacturing method of this invention, it turned out that the drug efficacy of the earthworm dry powder obtained by Japanese Patent Application Laid-open No. 59-216572 declines rapidly except the antidiabetic agent. Vacuum drying of the final finishing process was performed under the same conditions as the cavity materials. That is, according to pharmacological test 1, which describes the hypoglycemic test for experimental diabetic mice of the product earthworm dry powder obtained by varying the heating time to 20, 24, 26 and 30 hours at a temperature of 80 ° C. and a vacuum of 0.3 torr. Measured. Compared with the earthworm dry powder of the manufacturing method of this invention, it turned out that the efficacy is rapidly lost by about 47, 76, 81 and 89%, respectively.

Recently, according to one of the inventors of the present invention, such as Mihara Hisashi and other co-researchers, the earthworm active substance has a suitable pH of 8 to 10, a stable pH of 5 to 10, trajirol (trade name), transamine (trade name), It has been confirmed that it is an enzyme protein that is inhibited by soybean trypsin activator and serum, has a plasminogen activating action and a dermatin soluble action, and has an agent having no fibrinogen soluble action.

Japanese Patent Application Laid-Open Publication No. 58-148824 (Application Date 1982) (Patent No. 58-148824) for the preparation of a preparation enzyme protein fraction by extraction of an aqueous solvent from an earthworm and a preparation method for the active substance for use as a glycoprotein protein fraction by the purification thereof. February 27) and US Patent Application No. 470394 (filed February 28, 1983) of a foreign application pursuant to its priority, British Patent Application No. 8305359 (filed February 25, 1983), Italy 47795A (February 25, 1983), French Patent Application No. 03165 (February 25, 1983), West German Patent Application No. P3306944. No. 1 (filed February 28, 1983) and Canadian Patent Application No. 422034 (filed February 21, 1983) are reported.

In addition, Mihara Hisashi has applied for six new protease material patent applications from earthworms, that is, Japanese Patent Application Publication No. 59-63184 (filed October 2, 1982) and their proteases as active ingredients. Japanese Patent Application Publication No. 59-184131 (filed March 31, 1983) and a foreign patent application claiming priority of a combined application thereof, that is, Korean Patent Application No. 2990 (Application Date) June 30, 1983), including the following patent applications: [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 2, 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 result of the present inventors, the literature which reported the activity of the antidiabetic treatment, prevention agent, or hypoglycemic activity which uses the earthworm dry powder and earthworm extract as an active ingredient was not found.

In other words, no literature was reported that significantly lowered blood glucose levels when the dry powder of the earthworm and the extract of the earthworm were administered to experimental diabetic mice by alloxane.

In addition, human diabetic patients were given an earthworm dry powder for 4-9 months with meal therapy. In the case of patients with mild to moderate diabetes, the blood glucose level improved from 2 to 3 months after the administration, and after 4 months, the literature reported excellent effects such as lowering the blood sugar to the standard value of healthy people. I could not.

Moreover, the literature reporting the manufacturing method of the dry powder of an earthworm and the extract of an earthworm which has such an effect was not found.

Conventionally, as an oral antidiabetic agent, organic synthetic compounds, such as sulfonyl urea and biguaMde compounds, are widely used.

The present inventors have intensively studied for many years the use of earthworm dry powder for the purpose of the invention of a high-medicinal product which is not among such synthetic organic compounds and has no side effects and high safety in natural products. Surprisingly, the inventors have found that dry powders of earthworms and extracts of earthworms are effective in treating and preventing diabetes.

The present invention is a dry powder of the earthworm containing the dried product of the earthworm in some form; Regarding diabetes treatment and prevention agent containing earthworm extract obtained by extracting and treating the earthworm as an active ingredient obtained by extracting and treating the earthworm with at least one extractant selected from the group consisting of an aqueous solvent, a water miscible organic solvent and a water immiscible organic solvent. will be. In addition, the novel and progressive improvement methods shown below have been studied in detail to obtain a high yield of dry powder of aseptic earthworms, which have a safe and excellent therapeutic effect and can be stored or stored for at least 4 years in a closed state. Was established.

Preparation 1:

1 part (weight) of mountain worms in 1 to 10 parts (capacity) of fresh water or organic acids such as acetic acid, citric acid, succinic acid, malic acid, tartaric acid, lactic acid or inorganic acids such as phosphoric acid, sulfuric acid and hydrochloric acid, or sodium or potassium salts of these acids. Low concentration or PH 3 to 6. containing at least one compound of 0.3% by weight or less, preferably 0.1% by weight or less. In 1-10 parts (capacity) of 5 weakly acidic aqueous solution, it is left at a temperature of 1-25 degreeC for 0.5 to 72 hours, Preferably the temperature is 2-15 degreeC and 1 to 40 hours, The soil in the digestive tract of the mountain earthworm was sufficiently excreted. Then, the dirt attached to the body surface of the mountain earthworm was washed out with water, and then the water was removed and the earthworm taken out was ground with a grinder. 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 lyophilized and vacuum dried. The conditions of freeze drying and vacuum drying are 5 to 100 hours, preferably 10, while raising the temperature stepwise with a vacuum degree of -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 mountain worms after washing off dirt attached to the surface of the body with water for 10 to 30 hours at a temperature of 1 to 30 ° C, preferably 10 to 30 hours at a temperature of 4 to 28 ° C, and 1 to 10 parts of fresh water Or (at least) one or more compounds of organic acids such as acetic acid, citric acid, succinic acid, malic acid, tartaric acid, lactic acid or inorganic acids such as phosphoric acid, sulfuric acid and hydrochloric acid, or sodium or potassium salts of these acids. Or less, preferably a low concentration of 0.1 (% by weight) or less, or a pH of 3 to 6. Excretion power of the mountain worm itself by leaving 1 to 10 parts (capacity) of 5 to 5 to 72 hours at a temperature of 1 to 25 ° C., preferably 1 to 40 hours at a temperature of 2 to 15 ° C. The soil in the digestive tract of the mountain earthworm was sufficiently excreted, and then the earthworm taken out by washing with or without washing with water was crushed with a grinder. The thick suspension of the obtained earthworms was frozen at a low temperature below −5 ° C., preferably at −10 to −60 ° C., followed by freeze drying and vacuum drying. The conditions of the freeze-drying and vacuum drying are the temperature of -60 to +90 deg. C, the degree of vacuum of 100 mmHg or less, preferably of the degree of vacuum of -40 to +80 deg. Freeze-drying and vacuum drying were performed for 10 to 60 hours to obtain dry powder of aseptic earthworms.

As a method of 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, or a device of a pressurized cell destruction device to prepare a suspension or homogenate. The temperature at the time of this grinding | pulverization is 1-25 degreeC, Preferably 2-15 degreeC is preferable.

By the above operation, yellow-brown or brown earthworm dry powder was obtained from the mountain earthworm in a yield of 20 to 35%. In this case, the earthworm dry powder has 5 to 16% moisture, preferably 7 to 14% ash, 3 to 8% ash, preferably 4 to 7% nitrogen, and 1 to 11% nitrogen, preferably 6 to 6%. It was prepared to contain 11%. In the earthworm dry powder, 18 kinds of aspartic acid, threonine, serine, glutamic acid, proline, glycine, alanine, cysteine, valine, methionine, isoleucine, leucine, tyrosine, phenyalanine, tryptophan, lysine, histidine, arginine or after Contains amino acids

[Test Example 1]

Preparation of earthworm dry powder products (M-1 to M-6) obtained by the method of the above-mentioned manufacturing method 1 (late Example 17- Example 20) and manufacturing method 2 (late Example 21, 22, 24, and 25) The analysis results are shown in Table-1. The earthworm dry powder product symbol M-1 by the manufacturing method 1 was obtained by the method of Example 17. Similarly, M-2 was obtained in Example 18, M-3 was obtained in Example 19, and M-4 was obtained in Example 20. The earthworm dry powder product symbol M-5 by the manufacturing method 2 was obtained by the method of Example 21. Similarly, M-6 was obtained by the method of Example 22.

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 by the method of Preparation Method 1 and M-5 of the earthworm dry powder product obtained by the method of Preparation Method 2.

TABLE 2

[Test Example 3]

Amino acid analysis of crude protein in the earthworm dry powder product of M-2 obtained by the method 1 and the synthesis of M-4 and the method 2 was carried out, and the results of comparison with the analysis of the excess and soy meal of protein foods were compared. Table 3 is shown.

TABLE 3

* Crude proteins, lipids and various metals among the earthworm dry powders obtained by the methods 1 and 2 in Tables 2 and 3 in the data described in P. McDonald, RA Edwards and JFD Greenhalgh Animal Nutrition (1973). Was found to be abundant, and it was found that the amino acid composition in the crude protein contained a large amount of essential amino acids.

The present inventors remove dirt, such as mud, straw, rock salt, and clay attached to the body surface of mountain earthworms and clay in the digestive tract as soon as possible to improve the yield, physicochemical activity of the earthworm dry powder product, It was found to be an important operation for enhancing the preservation.

In particular, the method of the first method in which the soil in the digestive tract of the mountain earthworm was excreted first and then the dirt attached to the surface of the body was washed and removed with water was able to finish the cleaning operation in a shorter time than that of the second method.

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 dirt attached to the body surface before discharging the digestive tract, the earthworm excretes the soil in the digestive tract during its operation and attaches to the body surface of the earthworms. Not done. In order to clean, it is necessary to repeat washing 4 to 5 times as shown in Examples 15-19 later. As a result, there is a loss of earthworms and a loss of body fluids and digestive fluids from wounds of the living body with an increase in the number of cleaning operations.

However, in the case where the remaining soil is excreted after being left in the above aqueous solution or fresh water, there is a small amount of soil adhering to the body surface of the mountain earthworm. Therefore, when the amount is small, it may be good for light cleaning in the subsequent drying process, or when cleaning is unnecessary.

Yield and product analysis results of earthworm dry powder products obtained by the same type of red earthworms (Lumbricus rebellus) and using the same washing method as the manufacturing method 1 and the manufacturing method 2 but the subsequent manufacturing conditions are almost the same. A comparison of Table-1 and Table-2) is shown in Table-3-1-.

TABLE 3a

That is, as can be seen from the comparison between the products M-3 and M-5 of the products M-3 and M-5 shown in Table 3a, the earthworm dry powder product obtained by the method of Production Method 1 has a higher yield than the product of Production Method 2. 0.5% to 1%, ash is 0.7% to 0%. 9%, nitrogen is 0.4-1. 4% are all many. Moreover, the product of the manufacturing method 1 obtained the preferable product which tends to have 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, in the aqueous solution containing at least one compound of sodium or potassium salts of the above-mentioned organic acid or inorganic acid or a mass production thereof at a low concentration as described above, the earthworm is left in fresh water than that of the earthworm in the digestive tract of the earthworm. Excretion is fast and excretion rate is large.

In the manufacturing methods 1 and 2 of the former, the completion state of the earthworm dried product after completion | finish of drying is often a state in which it aggregated or the mass mixed. However, when this is crushed by a grinder, it becomes powder easily. When administering the dry matter of an earthworm and the extract of an earthworm to a mammal including humans, it is preferable that it is a powder rather than a lump.

Mihara Hisashis (Japanese Patent Application Laid-Open Nos. 59-63184 and 59-184131) fractionated six new proteases of a linear enzyme in earthworms. In other words, Mihara Hisashi added doubling physiological saline to earthworm dry powder for 2 days and incubated the supernatant for the supernatant. As a result of cellulose ion exchange chromatography, proteins of I, II, and III fractions having casein and dermatin degrading activities were obtained. The fractions of I, II, and III were purified again by DEAE-cellulose, Sephadex G-75, Toyopearl HW55, ACH-Sepharose, Benzamidine-Sepharose, and the like. Purified enzyme was obtained. When the molecular weight of SDS-PAGE was measured, the molecular weight of fraction I-0 was the lowest, and when it was calculated as 23, 500, the molecular weight increased sequentially to I-1, I-2, III-1, III-2, and III-2. Was the molecular weight 34, 200. The isoelectric point of the six fractions was measured by isoelectric point electrophoresis, with I-0 being the highest, pH 4. 12, and then the pH lowering sequentially and pH 3. 52 at III-2. These six fractions are new proteases that are different from the serine enzymes, and the most suitable pH of these six fractions is around 8 or pH 8-10, and the stable pH is 4-12 or 5-12, the most suitable. It is reported that temperature was 50 degreeC or 50-60 degreeC, and inactivation conditions were 70 degreeC for 60 minutes.

When 10 times the amount of physiological saline was added to the dry powder products M-4 and M-5 of the earthworms obtained in the above-mentioned preparation methods 1 and 2, the supernatant was measured by a standard dermatitis plate, and immediately showed the active activity as shown in Table-4. . Incubating the physiological saline solution of the earthworm dry powder of M-4 at 37 ° C. showed that the supernatant activity of the supernatant was about 4 times at 10 days and 5 times at 50 days and 5 days at 75 days as shown in Table-4. 5 times It is believed that this protease precursor is present in large amounts in the lyophilized powder of the earthworm, and there is an expression of enzymatic activity by self-extinguishing. Comparing the activity of the supernatant on the 50th day in terms of urokinase and other international units, it is calculated as about 8,000 IU ml. The enzyme also dissolves in the plasminogen free dermalin plate and the standard dermalin plate, and the standard dermalin plate has a larger solubility window compared to the plasminogen free plate and has the enzyme activity of directly decomposing dermatin. Plasminogen activator activity was also shown. In the above operation, the result of using the earthworm dry powder of M-5 instead of M-4, the same results as in Table-4 were obtained.

TABLE 4

By adding 10 times the amount of physiological saline to the earthworm dry powder of the present invention

Practical activity of the supernatant when incubated at 37 ° C

Although the earthworm dry powder and earthworm extract of the present invention are orally administered to rats and humans for the reason of showing glycemic effect or diabetes treatment and prevention effect, the details of protease contained in earthworm dry powder and earthworm extract are unknown. It is thought to be due to (protein) itself or its enzymes or precursors (proteins) of these enzymes themselves or proteins or lipids or other unknown compounds themselves or their combined action. The nitrogen content of the earthworm dry powder of the present invention most preferred as a therapeutic agent for diabetes is 7 to 10%, that is, crude protein content 43. 8 to 62. 5%. The nitrogen content of the earthworm extract of the present invention most preferred as a therapeutic agent for diabetes is 7-16%, that is, 43.8-100% crude protein content.

Next, preferred specific examples of the freeze-drying and vacuum-drying operations of the suspension by wet grinding of the earthworm obtained by the above-described methods 1 and 2 are as follows.

The ground mass of the earthworm, i.e., the suspension of the earthworm, is frozen at -10 to -60 DEG C, preferably at -30 to -50 DEG C for 5 to 10 hours, and then at 0.01 to 0 at the same temperature. Lyophilization was carried out under vacuum of 2 mmHg for 5 to 12 hours. Next, 0 to 01 to 0 for 5 to 15 hours at 20 to 30 ° C. Dry under vacuum of 2 mmHg.

Next, 35 to 50 ° C and 0.1 to 0. Drying under vacuum of 5 mmHg for 10 to 20 hours. The vacuum drying of the final finishing process is followed by drying for 20 hours. The vacuum drying of the next final finishing process is from 0.01 to 0. Vacuum drying at 5 to 10 hours at 70 to 80 ° C., preferably at 75 to 80 ° C., in a vacuum of 5 mmHg, yielded an aseptic and earthworm dry powder containing 5 to 15% of moisture. In particular, vacuum drying of the final finish is an important operation. In order to finish the powder of the earthworm in aseptic state without deactivating the activity of the protease and the precursor of the enzyme contained in the earthworm dry powder, the present inventors studied in detail the results of the lyophilization and vacuum drying. Vacuum degree of vacuum drying conditions 0.01-1. The combination of three elements of 5 mmHg, a heating temperature of 70-80 ° C. and a time of 5 to 10 hours was found to be an important condition and the above operating conditions were established.

As shown above, all six types of purified protease from the earthworm dry powder (Japanese Patent Application Laid-open Nos. 59-63184 and 59-184131) have been reported to deactivate at 70 ° C. for 60 minutes. Protease in the dry powder of the earthworm obtained by the production method of the invention is not inactivated as shown in Table-4. The reason is as follows.

In the method described in both Japanese Patent Application Laid-Open, six kinds of protease were inactivated by heating at 70 캜 for 60 minutes under normal pressure (1 atm). By the way, in the final process of the manufacturing method of the present invention, 0.01-0. Proteolytic enzymes in the earthworm dry powder do not deactivate due to heating at 70-80 ° C. for 5-10 hours under high vacuum of 5 mmHg. That is, a combination of the above-mentioned heating temperature, heating time, and vacuum degree as a preferable condition for vacuum drying in the final finishing process of the earthworm dry powder of the present invention becomes an important condition for sterilizing various bacteria without inactivating protease. These combination conditions are conditions established by the present inventors.

The earthworm dried powders obtained by the methods 1 and 2 were stored for 5 years in a sealed state at room temperature of 5 to 45 ° C. and no mold or other physical or chemical alterations were observed.

및

의 제법에 의해 수득한 지렁이의 건조물 및 그 건조분말이어도 좋다.The form of the earthworm dry powder other than the antidiabetic agent of the present invention is, in addition to the earthworm dry powder obtained in the above-mentioned production methods 1 and 2, conventionally known

And

The dry matter of the earthworm obtained by the manufacturing method of this, and its dry powder may be sufficient.

In addition, the extract of the earthworm other than the antidiabetic agent can be obtained by the following manufacturing method.

The earthworm of the raw material is extracted and treated with at least one extractant selected from the group consisting of an aqueous solvent, a water miscible organic solvent, and a water immiscible organic solvent, and the obtained extract is concentrated to precipitate as a precipitate or concentrated. An organic solvent was added to the extract to obtain a precipitate, or the extract was completely removed under normal pressure, pressure, or reduced pressure to obtain an extract.

및

의 공지의 제법에서 수득한 지렁이의 건조물 및 그 건조분말; 분토를 제거한 것 또는 분토를 함유한 산지렁이 또는, 이들의 날지렁이; 이들의 산지렁이 및ℓ또는 날지렁이의 분쇄현탁액 등의 형상의 지렁이를 사용할 수가 있다. 이러한 것 중, 가장 바람직한 원료 지렁이의 형태는, 상기한 제법 1 및 2의 조작처러 공정에 있어서 습식분쇄를 행하여서 수득한 지렁이의 현탁액이다. 즉, 산지렁이를 초산, 시트르산, 숙신산, 말산, 타르타르산 또는 젖산으로 이루어지는 유기산; 인산, 황산 또는 염산으로 이루어지는 무기산; 이들의 유기산나트륨염; 이들의 유기산칼륨염: 이들의 무기산나트륨염; 이들의 무기산칼륨염으로 이루어지는 군에서 선택된 적어도 1종류의 화합물은 0. 3(중량)% 이하의 저농도 또는 pH 3∼6. 5의 미산성의 수용액 중 또는 담수중에 방치해서 산지렁이의 소화관내의 분토를 토하게 한 후, 산지렁이의 체포면에 부착된 오물을 물로 세척 제거해서 습식 분쇄를 행하든가, 또는, 산지렁이의 체포면에 부착된 오물을 물로 세척 제거한 후의 산지렁이를 상기 수용액 중 또는 담수중에 방치해서 산지렁의 소화관내의 분토를 토하게 한 후, 습식분쇄를 행하여, 수득한 지렁이의 페이스트 상의 현탁액이다. 이 현탁액을 원료로 했을때에는, 다른 형태의 지렁이 보다도 용매의 사용량이 절약되고, 또한 당뇨병 치료제로서의 유효 물질을 단시간 내에 추출할 수가 있으며, 또 처리시간 단위 당의 추출물량이 많다는 등의 추출효울이 우수하다. 다음에 바람직한 원료형태는 상기 제법 1 및 2에서 수득한 지렁이 건조분말이다.Earthworms of the raw material, the earthworm dry powder obtained in the above-described production methods 1 and 2; As mentioned above,

And

Dried products of the earthworm obtained by the known production method of Dirt removed or mountain earthworms containing clay, or earthworms thereof; These earthworms and earthworms having a shape such as l or a groundworm may be used. Among these, the most preferable form of the raw earthworm is a suspension of the earthworms obtained by wet grinding in the above-described operation of steps 1 and 2. That is, the mountain earthworm is an organic acid consisting of acetic acid, citric acid, succinic acid, malic acid, tartaric acid or lactic acid; Inorganic acids composed of phosphoric acid, sulfuric acid or hydrochloric acid; Organic sodium salts thereof; Organic potassium salts thereof: their sodium salts; At least one compound selected from the group consisting of these inorganic potassium salts has a low concentration of 0.3% by weight or less or a pH of 3-6. Either the soil is allowed to vomit soil in the digestive tract of mountain worms in an acidic aqueous solution of 5 or in fresh water, and the soils attached to the surface of the mountain worms are washed and removed by water, or the earthworms are arrested. The earthworm after washing and removing the dirt attached to the cotton with water is left in the above aqueous solution or in fresh water to vomit soil in the digestive tract of the mountain earthworm, and then wet grinding is performed to obtain a paste-like suspension of the earthworm obtained. When the suspension is used as a raw material, the amount of solvent used is reduced compared to other types of earthworms, and an effective substance as an antidiabetic agent can be extracted within a short time, and the extraction effect such as a large amount of extract per treatment time unit is excellent. Next, the preferred raw material form is the earthworm dry powder obtained in the above production methods 1 and 2.

As the aqueous solvent of the extracting solvent, it is preferable to use a starting solution, distilled water, physiological saline, various buffers having a pH of 5 to 10, or various prepared waters. As the buffer, one or more of buffers having various compositions of pH 5 to 10, preferably PH 6 to 8, such as phosphoric acid, acetic acid, boric acid, citric acid, tris, and hydrochloric acid may be used. In addition, pH 5-10, Preferably the preparation liquid of pH 6-8 is water-soluble inorganic acid, such as hydrochloric acid, sulfuric acid, phosphoric acid, acetic acid, lactic acid, citric acid, succinic acid, malic acid, or tartaric acid, an organic acid, alkali metals, such as sodium and potassium It means the lean aqueous solution prepared by the hydroxide or carbonate of this. The most preferred aqueous solvents are physiological saline, buffers, and water. When using such an aqueous solvent, preservatives, such as parahydroxy benzoic acid ethyl ester, a sodium benzoate salt, and sodium adiide, are 0.01-0. It is preferable to use 3 (weight)%.

As a water miscible organic solvent, For example, C1-C4 lower alcohols, such as methyl, ethyl, allyl n-propyl, i-propyl, n-butyl, i-butyl, s-butyl, or t-butyl alcohol; Ketones having 1 to 4 carbon atoms such as acetone or methyl ethyl ketone; Ethers having 1 to 4 carbon atoms such as ethyl ether, 1, 2-propylene oxide, dioxolane, 4-methyldioxolane or dioxane; It is preferable to use C1-C4 esters, such as methyl formate, ethyl formate, and methyl acetate. Of these, the most preferable water miscible organic solvents are alcohols having 1 to 4 carbon atoms.

Examples of the water immiscible organic solvents include 6 to 10 carbon atoms such as n-hexane, i-hexane, n-heptane, i-heptane, n-octane, i-octane, cyclohexane, bengen, toluene, xylene or ethylbenzene. Hydrocarbons; 6 carbon atoms such as n-hexyl, 2-methylpentyl, 2-ethylbutyl, s-hexyl, s-heptyl, n-octyl, 2-ethylhexyl, 2, 6-dimethyl-4-heptyl, n-decyl alcohol, and the like Alcohol of -10; Ketones having 7 to 8 carbon atoms such as methyl-n-amyl ketone, methylcyclohexa or methylhexyl ketone; Esters having 6 to 8 carbon atoms such as s-butyl acetate, i-butyl acetate, 2-ethylbutyl acetate, s-amyl acetate or methyl amyl acetate; It is preferable to use halogenated hydrocarbons having 1 to 5 carbon atoms such as chloroform, dichloropentane or epichlorohydrin.

The extraction method may be performed by a general method and is not particularly limited, but one example thereof is as follows.

Aqueous solvent, concentration of 5 to 70 (volume)%, preferably aqueous solution of organic solvent of 10 to 60% (volume)%, water miscible organic solvent of concentration of 80 to 100 (volume)% 1-100 times (weight L volume) preferably at least 2-30 times at least 1 type of extractant chosen from the group which consists of a water-immiscible organic solvent of about 100% of concentration, and extraction temperature is -50- + 60 DEG C is preferably -40 to +60 DEG C, and the extraction contact time is from 10 minutes to about 100 days, preferably from 30 minutes to about 40 days, each time or continuously stirred, and then left to stand and filtered to extract an extract. . Filtration may use a pressurized or reduced pressure filter (group) or a centrifuge. The extraction residue is washed well and then the extract and wash are combined. The extract or mixed solution was concentrated under normal pressure, pressure, or reduced pressure at a temperature of 60 ° C. or lower, and the extraction solvent was removed by a concentration method such as ultrafiltration concentration or lyophilization to obtain an extract. However, when ethanol is used as the extractant, for example, ethanol may be left without being completely removed depending on the type of the final target formulation. Moreover, the crystal precipitation method by leaving the concentrate of the extract or mixed liquid mentioned above, or the method of obtaining an extract by filtering and drying after adding an organic solvent to precipitate a crystal | crystallization, and obtaining a crystal | crystallization is also used.

The yield of the extract with respect to the raw earthworm in the above operation is 3 to 60% by weight in dry matter. The difference in yield depends on the extraction conditions such as the shape of the raw earthworm, the type of solvent, the extraction temperature, and the extraction time. If the extraction temperature is high, the extraction temperature is preferably -40 to +60 ° C because there is a risk of decomposition of the effective substance.

As shown in Table-2 and Table-3, the components contained in the extract contain 18 kinds of amino acids, 8 kinds of metals and the like as the components of the dry powder of the earthworm. In particular, the protein in this extract contains a protein consisting of amino acids such as a raw earthworm, a saccharide protein, a metal binding protein and the like. The extract also contains free amino acids, lipids, various sugars, ATP and ARP-like substances, and other chemical compositions of unknown composition.

The earthworms used in the present invention are red earthworms (Lumbricus rubellus), LT earthworms (alias, fishing earthworms (Lumbricus terrestris),] spotted earthworms (Eisenia foetida), brown fishing earthworms (Allolobophora caliginosa), purple fishing earthworms (Dendrobaena octaedra), Alliumboworma (Allolobophora japonica Michaelsen), Haida worm (Drawida hatta mimizu Hatai), black earthworm (pheretima divergens Michaelsen), common earthworm (phertima commuMssima), fieldworm (phertima agretis), ciet bolt earthworm Common worms, such as pheretinn hilgendorfi, worm earthworms (pontodrilus matsushimensis Iizuka), earthworms (Tubifex hattai Nomura), and wallworm earthworms (aka lily earthworms) (Limnodrilus gotoi Hatai = L. Socialisstephenson). Ramen can use any worm.

When administering the earthworm dry powder and earthworm extract of the present invention for clinical treatment may be used alone or in combination thereof. The form may be any other than oral or parenteral, but oral administration is particularly preferable. As an oral dosage form of the present invention, a mixture of the present invention itself or a suitable pharmaceutically acceptable pharmaceutical carrier is used, such as capsules, tablets, granules, powders (powders), coating agents, dragees, emulsions and the like. Examples of the pharmaceutical carrier include, for example, lactose, white sugar, mannitol, glucose, starch, sorbitol, glycine, calcium phosphate, microcrystalline cellulose, and the like as excipients; As binder, starch, gelatin, gum arabic, glucose, sucrose, sorbitol, mannitol, tragant, hydroxy propyl cellulose, hydroxypropoxymethyl cellulose, carboxymethyl cellulose, 2, -methyl-5-vinyl Pyridine-methacrylic acid-methyl acrylate copolymer, polyvinylpyrrolidone, sodium alginate 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; Starch containing potato powder, surfactant, etc. as a disintegrating agent; Sodium lauryl sulfate etc. are mentioned as a humectant. In addition, when administered parenterally, it can be used as a suppository. In particular, cacao oil, witepsol, savanal, polyethylene glycol, polypropylene glycol, glycerogelatin, gelatin capsules and the like are used as the base material of the suppository. In addition, known safe preservatives, such as methyl para hydroxy benzoate, propyl para hydroxy benzoate, butyl para hydroxy benzoate, and butyl hydroxy anisole, and other safe pigments are used in combination.

The dosage of the earthworm dry powder and the earthworm extract in addition to the anti-diabetic agent of the present invention also varies depending on the administration method, age, weight, condition, and type of disease of the patient, but it is generally preferred for a person from about 0.01 g to about 5 g per day. . Most preferably, it is divided into 1 to 3 times a day from 0.2g to 2g per day.

The toxicological and diabetic therapeutic effect of the earthworm dry powder of the present invention or the pharmacological test method of blood sugar drop and the results thereof will be described in detail below.

[A. Acute Toxicity Test:]

An acute toxicity test was performed in oral administration using 5 ddy male mice weighing 30 ± 2 g and 5 wistar male mice each weighing 100 ± 2 g. Earthworm dry powder M-1 of the present invention (water 10. 2%, ash 5. 1%, nitrogen 9. 5%); Copper M-2 (containing 10. 4% moisture, 5. 3% ash, 8. 4% nitrogen); Copper M-3 (contains 10. 7% moisture, 5.2% lean, 9.2% nitrogen); Copper M-5 (containing 10.6% moisture, 5.6% ash, 9.6% nitrogen); Copper M-5 (contains 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 added from 0.19 L kg to 8 g L kg, to the mice (0.1 to 5 g L kg) and rats described above. (2 to 8 gL kg) were individually dosed by forced administration with a throat scrubber bar. Animals were maintained at 22-23 ° C. during the test period and observed for 14 days after dosing. No deaths as doses were seen. Post-dose intoxication and behavior were observed over time, but there was no difference between normal animals and lower animals. In addition, weight gain was almost no difference from normal animals. There was no macroscopic obstacle in any part of the main organs in the post-mortem examination. Therefore, the earthworm dry powder of the present invention could not determine the LD ₅₀ value because it is very low toxicity.

[B. Actions on Synthetic Diabetic Mice]

[Pharmacology Test 1]

1. Animals: One group of ddy male mice weighing 30 ± 2 g and 5 mice were used.

2. Feed and breeding conditions: Solid feed made by Japan Cree Co., Ltd. was used. One mouse was placed in one cage, and the feed and water were ingested freely. It was raised at a constant temperature and humidity of 23 ± 1 ° C. and 55 ± 5% of humidity.

The ddy male mice were fasted for 16 hours, followed by intravenous administration of 75 mg lkg of oxalate, 48 hours after the dry powder M-1 of the present invention (water 10. 2%, ash 5. 1%, nitrogen 9. 4%), Copper M-2 (Moisture 10.4%, Ash 5.3%, Nitrogen 8.6%), Copper M-3 (Moisture 10.7%, Ash 5.2%, Nitrogen 9.2%), Copper M -4 (moisture 10.6%, ash 5.6%, nitrogen 9.6%), copper M-5 (moisture 9.5%, ash 4.5%, nitrogen 7.8%), copper M-6 Six kinds (300mgℓkg) of (moisture 9.2%, ash 4.7%, nitrogen 8.4%) were suspended orally in saline solution, respectively, and blood was collected from the heart after 150 minutes, and blood glucose equivalent was measured by glucose oxidase method. Was measured.

The measurement results are shown in Table-5. Earthworm dry powder of this footing was found to significantly lower blood sugar.

TABLE 5

*: P <0 L05, * * P: <0. 01, * * *: P <0. 001

[Action- II:]

The pharmacological test method and the results of the toxicity and hypoglycemic drop of the earthworm extract of the present invention will be described below.

[Preparation of Raw Material:]

[Manufacturing 3:]

[Preparation of Earthworm Dry Powder:]

중에 온도 15℃에서 3시간 방치하여, 소화관내의 분토를 충분히 배설시켰다. 이 산지렁이를 물로 잘 세정하여 산 지렁이의 체표면에 부착되어 있는 진흙, 분 등의 오물을 씻어낸 후, 간단히 물끼를 빼고 꺼낸 산지렁이 그대로를 울트라호모믹서(닛뽄 세이기 주식회사 제)로 분쇄하여, 지렁이의 진한 현탁액 9. 3kg을 수득하였다. 이 진한 지렁이 현탁액을 1. 55kg씩으로 6등분하여, 각각을 E-1, -2, -3, -4, -5 및 -6으로 하였다.Acidic aqueous solution of pH 5.8 in which 10 kg (approximately 20,000) of mountain earthworms (red earthworms) are dissolved and a mixed acid of malic acid and lactic acid 2: 1 is dissolved.

The mixture was left to stand at a temperature of 15 ° C. for 3 hours to sufficiently excrete the soil in the digestive tract. After washing the mountain earthworm well with water, wash off the dirt, such as mud, powder, etc. attached to the body surface of the mountain earthworm, and simply remove the water and remove the earthworm as it is. 9. 3 kg of a thick suspension of earthworms was obtained. The thick earthworm suspension was divided into six equal parts of 1.55 kg each to E-1, -2, -3, -4, -5 and -6.

A thick suspension of the earthworms of E-1 and E-2 obtained above was placed in each tray and frozen at −40 ° C. for 30 hours. The product temperature was then lyophilized by vacuum at 0.1 mmHg at −40 ° C. for 6 hours, then dried at 25 ° C. under vacuum of 0.1 mm Hg for 8 hours, and then 12 hours under vacuum of 0.1 mm Hg at 45 ° C. 318 g of an earthworm's dried product was obtained in each of the two trays by drying and finally drying at 80 ° C. under vacuum of 0.1 mmHg for 6 hours. This was set to K-1 and K-2.

[Preparation of Extracts:]

X-①. 5 times 99% methanol 1.575L was added to 315 g of the earthworm dried product (K-1), and extracted three times at 25 ° C. Finally, the residue was washed with 0.95 L of three times 99% methanol. It was concentrated under reduced pressure at 30 ° C., concentrated to dryness and then vacuum dried to give 88.2 g of powder extract. This was used as extraction wool X-①.

X-②. To 315 g of the earthworm's dry matter (K-2), a mixed solution of 0.34 l of chloroform and 2.63 l of ethanol was added two times, and extracted three times at 20 ° C. The mixture was washed with 63 L, and the mixed liquid of the extract and the washing liquid was concentrated to dryness at 30 deg. C under reduced pressure, and then vacuum dried to give 72. 5 g of powder extract. This was taken as extract X-2.

를 수득하였다. 이 혼합액 7. 7

에 n-헥산 2. 31l를 가해서 교반한 후, 정치하여, 수용부층과 n-헥산층으로 분리후, 수용부층을 분취하여, 30℃에서 감압 농축 건조한 후, 동결건조하여 분말 추출물 86. 8g을 수득하였다. 이것을 추출물 X-③로 하였다.X-③. To 1.55 kg of the earthworm suspension (E-3), fresh water 6. 2 is added, stirred at 10 ° C. for 2 hours, filtered and separated into an extract and a residue. The residue was washed with water and then mixed with the extract and the wash solution. 7. 7

Obtained. This mixture 7.7

2.31 l of n-hexane was added to the mixture, followed by stirring. After standing, the mixture was separated into an aqueous layer and an n-hexane layer. The aqueous layer was separated, concentrated under reduced pressure at 30 deg. C, and lyophilized to obtain a powder extract. Obtained. This was taken as extract X-③.

로 세척한 후 추출액과 세척액과의 혼합액을 40℃에서 감압 농축 건조한 후, 진공건조하여, 분말의 추출물 69. 7g을 수득하였다. 이것을 추출물 X-④로 하였다.X-④. Earthworm Suspension (E-4) 1. To 55 g, 10 L of 40% ethanol aqueous solution is added, stirred at 40 DEG C for 12 hours, filtered and the mixture is separated into an extract and a residue. The residue is 40% ethanol 0.3

After washing with water, the mixed solution of the extract and the washing solution was concentrated under reduced pressure at 40 ° C and dried, and dried in vacuo to give 69. 7 g of an extract of a powder. This was taken as extract X-④.

를 수득하였다. 이 혼합액에 아지드화나트륨 19을 가하여 3℃에서 8시간 인큐베이션한 후, 30

에서 감압농축하여 수득한 농축액에 2배량의 에탄올을가하고 침전물을 여과하여 분취한 침전을 진공건조시키고, 다음에 동결건조시켜서 분말의 추출물 64. 2g을 수득하였다. 이것을 추출물 X-⑤로 하였다.X-⑤. Earthworm Suspension (E-5) 1. To 55 kg, pH 6.4 phosphate buffer solution 2. 5 was added and incubated at 37 ° C. for 8 hours to promote the action of the protease originally contained in the earthworm body, followed by filtration. Was washed with a pH 6. 4 phosphate buffer solution and the mixture was extracted and washed 3.9

Obtained. Sodium azide 19 was added to this mixed solution and incubated at 3 ° C for 8 hours, and then 30

To the concentrated solution obtained by concentrating under reduced pressure at 2 times, ethanol was added twice, and the precipitate was filtered off and the precipitate was collected by vacuum drying and then lyophilized to give 64.2 g of an extract of the powder. This was taken as extract X-⑤.

X-⑥. Earthworm Suspension (E-6) 1. After 55 kg of natural drying in sunlight, 900 ml of a mixed solvent of ethanol and ethyl ether 2: 1 was added to a dry powder of powdered earthworms (K-3). Extracted three times. Finally, the residue was washed with the mixed solvent described above, and the mixed solution of the extract and the washed solution was concentrated to dryness under reduced pressure at 30 ° C, and then vacuum dried to give 61. 5 g of an extract. This was taken as extract X-⑥.

[Manufacturing 4:]

[Preparation of Earthworm Dry Powder:]

중에 온도 8℃에서 3시간 방치하여, 소화관 내외 분토를 충분히 배설시킨 후, 산지렁이를 물로 잘 세척해서 산지렁이의 체표면에 부착되어 있는 진흙, 분 등의 오물을 씻어낸 후 간단히 물기를 빼고 꺼낸 산지렁이 그대로를 믹서에 걸어서 분쇄하여 지렁이의 진한 현탁액 9. 0kg을 수득하였다. 이 지렁이의 진한 현탁액을 1. 5kg씩으로 6등분하여, 그것을 E-7, -8, -9, -10, -11및 -12로 하였다. 이들 중 E-7과 E-8의 지렁이의 진한 현탁액 1. 5kg을 각각의 트레이에 넣고, -30℃에서 40시간 동결한 후, 다음에 품온 -40℃에서 0. 1mmHg의 진공하 6시간 동결건조시키고, 다음에 트레이가 실려있는 선반온도를 3℃로 올려 0. 1mmHg의 진공하에서 6시간 건조시킨 후, 선반온도를 50℃로 올려 0. 2mmHg의 진공하에서 8시간 진공건조시켜, 최후로 선반온도를 78℃로 올리고, 0. 2mmHg의 진공하에서 8시간 건조시킴으로써 지렁이의 건조분말 305g씩을 수득하였다. 이것을 K-7과 K-8로 하였다.Farmed earthworms (red earthworms) 13 kg (approximately 26,000 animals) of soil attached to the surface of the soil (bunto, dark black, straw, etc.) after washing with water, PH dissolving the mixed acid of malic acid and citric acid 1: 1 6. Acidic aqueous solution of 2 40

After leaving for 3 hours at a temperature of 8 ° C, the soil inside and outside the digestive tract is sufficiently excreted. After washing the earthworms well with water, the dirts, etc. attached to the body surface of the earthworms are washed out. The mountain earthworms were ground on a mixer and ground to obtain 9.0 kg of a thick suspension of earthworms. The thick suspension of the earthworms was divided into 1.5 pieces by 1.5 kg, which was E-7, -8, -9, -10, -11 and -12. Of these, 1. 5 kg of a thick suspension of the earthworms of E-7 and E-8 were put in each tray, and frozen at -30 ° C for 40 hours, and then frozen at -40 ° C for 6 hours under vacuum of 0.1 mmHg. After drying, the shelf temperature on which the tray is loaded is increased to 3 ° C. and dried for 6 hours under a vacuum of 0.1 mmHg. Then, the shelf temperature is increased to 50 ° C. for 8 hours under vacuum of 0.2 mm Hg, and finally the shelf is dried. The temperature was raised to 78 ° C. and dried for 8 hours under a vacuum of 0.2 mmHg to obtain 305 g of dry powder of the earthworm. This was called K-7 and K-8.

[Preparation of Extracts:]

에 0. 02% 파라이드록시벤조산 에틸에스테르를 함유하는 0. 9% 염화나트륨 수용액을 가하여, 30℃에서 96시간 교반시킨 후, 여과하여 잔사를 0. 9

의 0. 9% 염화나트륨 수용액으로 세척하고 추출액과 세척액을 합한 청정 추출액 3. 8

을 수득하였다. 이것을 한외여과법으로 농축하여, 액량을 0. 16

로 하고, 여기에 에탄올 0. 16

를 가해서 침전분별한 후의 여과액에 최종 농도로서 에탄올 농도로서 에탄올 농도가 80%가 되도록 첨가해서 수득한 침전을 진공 건조시켜서 결정분말 14. 4g을수득하였다. 이것을 추출물 X-⑦-1로 하였다.X-⑦. It is 3 for 300 g of earthworm dry matter (K-7)

To a 0.9% aqueous sodium chloride solution containing 0.02% of parideoxybenzoic acid ethyl ester was added to the resulting mixture, the mixture was stirred at 30 ° C for 96 hours, and then the residue was filtered.

Washed with an aqueous 0. 9% aqueous sodium chloride solution and extract and wash solution combined 3. 8

Obtained. This was concentrated by ultrafiltration and the liquid amount was 0. 16.

And ethanol here.

The precipitate obtained by addition was added to the filtrate after fractionation to obtain an ethanol concentration of 80% as a final concentration as a final concentration. The precipitate obtained was vacuum dried to obtain 14. 4 g of crystal powder. This was taken as extract X-⑦-1.

72. g of crystalline powder was obtained by concentrating and drying the filtrate of the extract X- 7-1 at 30 deg. This was taken as extract X-⑦-2.

를 수득하였다. 이 추출액을 한외여과법으로 농축한 액을, 다음에 감압 농축건조함으로써 69g의 결정분말을 수득하였다. 이것을 추출물 X-⑧로 하였다.X-⑧. To 300 g of earthworm dry matter (K-8) was added fresh water containing 3 l of 0.01% sodium benzoate, stirred and extracted at 32 ° C. for 72 hours, and then filtered and the residue was washed with fresh water of 0.6. Combined Clean Solution 35

Obtained. The liquid obtained by concentrating this extract by ultrafiltration was then concentrated under reduced pressure to give 69 g of crystal powder. This was taken as extract X-⑧.

의 0. 02%의 파라히드록시 벤조산 에틸에스테르를 함유하는 증류수를 가하여, 32℃에서 120시간 교반한 후, 여과하여 추출액과 잔사로 분취한다. 잔사를 증류수로 잘 세척한 후, 추출액과 세척액의 혼합액을 30℃에서 약 110양으로 강압 농축한 후, 4배량의 에탄올을 가하여 잘 교반한 후 1주야 정치하여 생성된 침전을 여과하여 에탄올로서 세척한 후, 진공건조시켜 분말추출물 72. 5g을 수득하였다. 이것을 추출물 X-⑨로 한다.X-⑨. Earthworm Suspension (E-9) 1. 5 kg was vacuum-dried at 40 ° C, and 3 to dry powder of powdered earthworm (K-9).

Distilled water containing 0.02% of parahydroxy benzoic acid ethyl ester was added thereto, stirred at 32 ° C. for 120 hours, and filtered and separated into an extract and a residue. The residue was washed well with distilled water, and the mixture of the extract and the wash solution was concentrated under pressure to about 110 at 30 ° C., followed by stirring with 4 times the amount of ethanol, and the mixture was left to stand overnight and the resulting precipitate was filtered and washed with ethanol. After drying in vacuo, 72. 5 g of powder extract was obtained. Let this be extract X-⑨.

를 가하여, 밀전 상태에서 때때로 교반하면서 20∼22℃에서 8일간 정치하였다. 그후, 여과하여 잔사를 50% 에탄올 수용액 0. 5로 세정하여 추출액과 세정액의 혼합액을 30℃에서 감압 농축 건조한 후 동결건조시켜 분말 추출물 40. 5g을 수득하였다. 이것을 추출물 X-⑩으로 하였다.X-⑩. Earthworm Suspension (E-10) 1. 60% ethanol in 5kg 7. 5

Was added and allowed to stand at 20 to 22 ° C. for 8 days with occasional stirring in a closed state. Thereafter, the residue was filtered, the residue was washed with 50% aqueous 50% ethanol solution, and the mixed solution of the extract and the washing solution was concentrated under reduced pressure at 30 ° C, dried and lyophilized to obtain 40. 5 g of a powder extract. This was taken as extract X-VIII.

와 50% 이소프로잔을 수용액 3

의 혼합 용매를 가하여, 밀전 상태에서 때때로 교반하면서, 8℃에서 16일간 정치하였다. 그후, 여과하여 잔사를 50% 에탄올 수용액 0. 5

로 세척하여, 추출액과 세척액의 혼합액을 35℃에서 감압 농축 건조한 후, 동결건조시켜 분말 추출물 28. 2g을 수득하였다. 이것을 추출물 X-⑪로 하였다.Suspension of X-worm earthworm (E-11) 1. 50% ethanol in 5 kg 4. 5

50% isoprozanol with aqueous solution 3

The mixed solvent of was added, and it left still at 16 degreeC for 16 days, stirring occasionally in a tight state. Thereafter, the residue was filtered to obtain 50% ethanol aqueous solution 0.5.

After washing with water, the mixed solution of the extract and the washing solution was concentrated under reduced pressure at 35 ° C and dried, and lyophilized to give 28. 2 g of a powder extract. This was taken as extract X-VIII.

를 가하여, 25℃에서 2시간 교반하면서 3회 추출하여 최후로 잔사를 60% 에탄올 수용액 1

로 세정하여 추출액과 세정액의 혼합액을 30℃에서 감압 농축건조하여, 다음에 동결건조함으로써 90. 5g의 분말 추출물을 수득하였다. 이것을 추출물 X-⑫)로하였다.X-⑫. Earthworm suspension (E-12) 1. 60% ethanol in 5 kg 7. 5

Was added and extracted three times with stirring for 2 hours at 25 ° C.

The mixture was washed with water, and the mixed solution of the extract and the washing solution was concentrated to dryness at 30 deg. This was taken as extract X-IX).

Toxicity and blood sugar of the extract obtained by extracting the earthworm extract, ie, the raw earthworm, by extracting and treating the raw earthworm with at least one extractant selected from the group consisting of an aqueous solvent, a water miscible organic solvent, and a water immiscible organic solvent. The pharmacological test method of lowering activity and the result are as follows.

A-1. Acute Toxicity Test:]

Electrical A. It was measured by the same method as the acute toxicity test. That is, an acute toxicity test in oral administration was performed using 5 ddy male mice each having a body weight of 30 ± 2 g.

For example, the above-mentioned extracts of the earthworms of the present invention, X-①, -②, -③, -④, -⑥, -⑨, -⑩ and -⑫, using the eight species, the mouse from 0.1 to 59 lkg And administered individually by forced administration to the throat probe rods. During the test, the mice were kept at an animal room temperature of 22 to 23 ° C and observed for 14 days after administration. Death at the administered dose was not entirely recognized. In addition, abnormal findings or signs of intoxication were not recognized. Therefore, the aqueous solvent and the extract by L or organic solvent from the earthworm of the present invention could not determine the LD ₅₀ value because of very low toxicity.

[Pharmacological examination 2:]

It was measured by the same method as the above pharmacological test 1 (Test of the effects of the action on experimental diabetic mice).

1. Animals: Five groups of 1 ddy male mice weighing 30 ± 2 g were used.

2. Feed and breeding conditions: Using a Nippon creased solid feed, one mouse was placed in one cage, and the feed and water were taken freely. It was raised at a constant temperature and humidity of 23 ± 1 ° C. and 55 ± 5% of humidity.

After fasting the above-mentioned ddy male mice for 16 hours, 75 mg of alloxane was administered intravenously, and 48 hours later, four kinds of earthworm dry powders of the present invention obtained in the above-described methods 3 and 4 [K-1,- 300 mg L kg of 3, -7 and-female;

Each 50mgℓkg of nine species [X-①, -②, -③, -④, -⑤, -⑥, -⑩, -⑪ and -⑫] of the earthworm extract of the present invention are suspended orally administered in physiological saline, respectively. After 150 minutes, blood was collected from the heart, and the blood equivalent was measured by the glucose oxidase method.

The measurement results are shown in Table-5-1. The dry powder and earthworm extract of the earthworm of the present invention was found to significantly lower blood sugar. The earthworm extract had a larger decrease in blood sugar level than the dry powder of the earthworm in the amount of 1 L16 of the earthworm dry powder.

Table 5a

*: P <0. 05 * *: P <0. 01 * * *: P <0. 001

[C. Oral Dose Experiments on Humans of Earthworm Dry Powders:]

Earthworm dry powder (M-3, water 10. 7%, ash content as described above in capsule C [1 capsule prepared in Example 6 to be described later along with meal therapy to five volunteers who obtained the test) 5. %, Nitrogen 9. 2%) 150mg] once capsules three times a day oral administration within 30 minutes after eating. Blood collection was done every month after the administration of only 62-year-old males. A 59 year old male was bled before and 2, 3 and 4 months after dosing. The 76-year-old woman was bled before and 4 months after dosing, and the 79-year-old woman was taken before and 3 and 6 months after dosing. A 61-year-old woman collected blood before and 1, 3, 4 and 8 months after dosing. Blood equivalents were measured by glucose oxidase preparation. The results are shown in Table-6.

TABLE 6

[Note] 9M ^a : 9 months later, the same meaning

The standard blood glucose level (mgℓdl) of local health workers who conducted this experiment was 50-100 before breakfast, 150 or less after 2 hours of breakfast, 50 to 100 before lunch, 150 or less after 2 hours of lunch, and 50 before dinner. It is less than 150 after two hours of dinner. The five volunteers shown in Table-6 also had high blood glucose levels and diabetes mellitus in all fasting blood predrugs before drug administration.

A 62-year-old man considered to be moderately diabetic showed an improvement in Hull equivalents 2 months after the earthworm dry powder administration of the present invention, and 3 months after the administration, 1 item after 2 hours of dinner. The value of 185mgℓdl was higher than the standard value. Four months after the administration, all six test values reached standard values. After that, 6 items of blood equivalents were maintained within the standard range for 5 months.

The 59-year-old male reached the standard value except that the value of 1 item before lunch was 114 mg Ldl among the 6 items after 3 months of administration, and all 6 items were within the standard values after 4 months of administration.

For 79-year-old women, the value of 4 items improved to within the standard range of 6 months after 3 months of administration, except for the slightly higher values of 171 and 107 mg Ldl at 2 hours after lunch and before dinner. After that, the standard value was completely reached.

The 76-year-old woman was mildly diabetic, but after 4 months of administration, the blood equivalents of 6 items reached standard values.

A 61-year-old woman is a very serious diabetic. After 4 months of administration of the earthworm dry powder of the present invention, the blood equivalents were improved. After 8 months, 4 items except for 2 hours after breakfast and before lunch were as high as 197 and 210 mg dl, respectively. The standard has been reached.

Oral administration of the dry powder of the earthworm to humans was safe and the side effects did not occur at all and could be safely terminated. For example, even in long-term administration for 6 to 9 months, there was no risk of hypoglycemia, which lowered the blood glucose level below the lower limit of the standard, and the earthworm dry powder of the present invention was found to be a safe drug.

The above results are summarized as follows. In the case of oral administration of the earthworm dry powder, it was slightly difficult to lower the blood equivalent of all 6 items to the standard value in the severely diabetic patients, but after 8 months, the blood equivalent of 4 of the 6 items was lowered to the standard value. I found that there is an improvement effect. However, sir. In patients with moderate diabetes mellitus, blood equivalents of all six items were lowered to within the standard level after 4 months after administration. Earthworm dry powder of the present invention is safe and excellent hypoglycemic agent, excellent diabetes treatment. I knew it was a preventive agent.

According to the above results, the earthworm extract, which is a solvent extract from the earthworm, is also safe for humans as well as an excellent blood sugar lowering agent and an excellent diabetes treatment in the same manner as the earthworm dry powder. We found out that it was preventive. That is, although there is a slight difference between the extract of the earthworm and the dry powder of the earthworm, it is considered to be a natural result since it is produced from a mountain earthworm or a raw earthworm of the same starting material.

Example 1

Tablet A

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, lactose, microcrystalline cellulose and corn starch of the earthworm were mixed well using a fluidized bed granulator according to the above formulation, and a 5% aqueous solution of hydroxypropyl cellulose was sprayed as a binder, followed by granulation after low temperature drying. .

Example 3

Granule B

Granule C

Granules were prepared by extruder mixing the well mixed powder according to the above formulation.

Example 4

Capsule A

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

Example 5

Capsule B

A hard capsule was prepared by filling 250 mg of the granule C prepared in Example 3 into the determined capsule.

Example 6

Capsule C

Mix the above prescribed well and mix this powder. 250 ml each was filled in a gelatin capsule of 1 to prepare a capsule.

Example 7

Enteric tablets

In accordance with the above-described prescription, the powder was uniformly mixed with a tableting machine to produce a predetermined amount, and then coated with the coating agent of the enteric coating film shown to prepare an enteric tablet.

Coating agent

Example 8

Powder A

Powder

Each of the above components was mixed well in a cone mixer to prepare a powder.

Example 9

Suppository A

Suppository B

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

Example 10

Capsule D

The prescription was mixed well. This mixed powder was No. 250 ml each was filled into 1 gelatin capsule, and the capsule was prepared.

Example 11

Tap Sulze E

In the above formulation, components other than hydroxypropyl cellulose were mixed well by using a fluidized bed granulator, and then a 5% aqueous solution of hydroxypropyl cellulose was sprayed as a binder and dried at 40 ° C. to obtain granules. The granules were filled in 250mg of light capsules to prepare light capsules.

Example 12

Capsule F

Granules

According to the above formulation, the well mixed powder was made into granules by an extruder. The granules were filled with 250 mg of seed capsules to prepare a capsule.

Example 13

Capsule G

The formulations described above were mixed well, and the mixed powder was packed into gelatin capsules by 150 mg to prepare capsules.

Example 14

Capsule H

The prescription was mixed well. This mixed powder was filled with 150 mg of gelatin capsures to prepare capsules.

Example 15

Capsule I

Mix well as prescribed above. 250 mg of each of the mixed powders was packed into capsules, thereby preparing a capsule.

Example 16

Capsule J

The prescription was mixed well. 250 mg of each of the mixed powders was filled into capsules, thereby preparing a capsule.

Example 17

Lightly washed with water acid worm (red worms) 1kg (about 2000), the malic acid and citric acid of 1: allowed to stand for 3 hours at temperature 18 ℃ the acidic aqueous solution of pH 2 to 6. The mixed acid dissolved in 1 l 4 and enteral After excretion of soil, the earthworms are washed well with water to wash away the dirt and dust attached to the body surface of the earthworms.

Next, it is pulverized by hanging on a mixer. The obtained suspension of the earthworms was put in a tray and frozen at -30 ° C for 40 hours, and then freeze-dried for 6 hours under vacuum of 0.1 mmHg at a temperature of -40 ° C, and then the temperature of the shelf on which the tray was loaded was raised to 30 ° C. After vacuum drying for 6 hours under vacuum of 0.1 mmHg, the shelf temperature was then raised to 50 ° C. and vacuum drying for 10 hours under vacuum of 0.2 mmHg, followed by raising the temperature of the shelf to 80 ° C. under vacuum of 0.2 mmHg. By drying for 8 hours, 280 g of dry powder product (M-1) of the earthworm was obtained.

Example 18

중에 온도 10℃에서 2. 5시간방치하여 소화관 내의 분토를 충분히 배설시킨후, 산지렁이를 물로 잘 세정하여 산지렁이의 체표면에 부착되어 있는 진흙, 분 등의 오물을 씻어 내었다.1 kg of acid worms (red worms) lightly washed with water, acidic aqueous solution of pH 5. 5 containing mixed acid of malic acid, tartaric acid and lactic acid 1: 1: 1

After leaving for 5 hours at a temperature of 10 ° C., the soil in the digestive tract was sufficiently excreted. Then, 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 obtained suspension of the earthworms was put in a tray and frozen at -25 ° C for 20 hours, and then the product temperature was lowered to -35 ° C and lyophilized for 7 hours under vacuum of 0.1 mmHg, and then the shelf temperature on which the tray was loaded was 28 ° C. The oligo dried powder product (M) was dried 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. -2) 275 g were obtained.

Example 19

1 kg of mountain worms (red worms) washed with water was left for 16 hours at a temperature of 10 ° C. in 3 L of fresh water to excrete the soil in the digestive tract. Wash the earthworms well with water to wash away the dirt and dust attached to the body surface of the earthworms. Then it was wet milled with an ultra homo mixer. The suspension of the obtained earthworms 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 a vacuum of 0.1 mm Hg, followed by drying for 10 hours under a vacuum of 0.08 mm Hg at 40 ° C., followed by a vacuum of 0.2 mm Hg at 40 ° C., 15 245 g of earthworm dried powder product (M-3) was obtained by drying time and then drying at 78 ° C. under vacuum of 0.1 mmHg for 8 hours.

Example 20

1 kg of a mountain worm (red worm) lightly washed with water was left in 2 l of an acidic aqueous solution of pH 5. 8 dissolved in malic acid at a temperature of 13 ° C. for 3 hours to sufficiently excrete the soil in the digestive tract. The mountain earthworms were washed well with water to wash away the dirt, such as mud and powder, which are 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 at a temperature of -30 ° C under vacuum of 0.1 mmHg for 8 hours, and then the temperature of the shelf on which the tray was loaded was raised to 25 ° C. Vacuum dry for 7 hours under vacuum of 0.1 mmHg, then lower the temperature of the shelf to 45 ° C., vacuum dry for 12 hours under the vacuum of 0.1 mmHg, and then raise the temperature of the shelf to 80 ° C. under vacuum of 0.1 mmHg. Earth drying to obtain earthworm dry powder product (M-4) 2759.

Example 21

중에 온도 15℃에서 18시간 방치하여 소화관내의 분토를 배설시켰다. 다음에 산지렁이를 가볍게 씻은 후 울트라 호모믹서로 습식 분쇄하였다, 수득된 지렁이의 현탁액을 트레이에 넣어서 -40℃에서 24시간 동결하였다. 다음에 품온을 -40℃에서 0. 1mmHg의 진공하에서 5시간 동결 건조시키고 다음에 25℃에서 0. 1mmHg의 진공하에서 8시간 건조시킨후, 다음에 45℃에서 0. 1mmHg의 진공하에서 12시간 건조시키고 다음에 80℃에서 0. 1mmHg의 진공하에서 7시간 건조시킴으로써 지렁이 건조 분말제품(M-5) 240g을 수득하였다.Dirt, straw, etc. adhering to the surface of 1 kg of mountain earthworms (red earthworms) were washed well with water five times and washed. Then take this mountain worm freshwater 2.5

It left for 18 hours at the temperature of 15 degreeC in the middle, and excreted the clay in a digestive tract. The earthworms were then washed lightly and wet-pulverized with an ultra homomixer. The suspension of the earthworms obtained was placed in a tray and frozen at −40 ° C. for 24 hours. The product temperature was then lyophilized for 5 hours under vacuum of 0.1 mmHg at −40 ° C., then for 8 hours under vacuum of 0.1 mmHg at 25 ° C., and then for 12 hours under vacuum of 0.1 mmHg at 45 ° C. 240 g of earthworm dried powder product (M-5) was obtained by making it dry for 7 hours at 80 degreeC under vacuum of 0.1 mmHg.

Example 22

Earthworms (red earthworms) 1 kg of soil, such as mud, powder attached to the surface of the body was washed well with water four times and washed.

중에 온도 15℃에서 2. 5시간 방치하여 소화관내의 분토를 배설시켰다. 다음에 산지렁이를 가볍게 세척한 후, 믹서를 습식 분쇄하였다. 수득된 지렁이의 현탁액을 트레이에 넣어 -35℃에서 24시간 동결시켰다. 다음에 품온 -35℃에서 0. 1mmHg의 진공하 7시간 동결 건조시키고 다음에 트레이가 실려있는 선반온도를 22℃로 올려서 0. 1mmHg의 진공하, 10시간 진공 건조시킨 후 다음에 선반의 온도를 42℃로 올려서 0. 2mmHg의 진공하 15시간 건조시키고, 최후로 선반의 온도를 78℃로 올려서 0. 1mmHg의 진공하에서 7시간 건조시킴으로써 지렁이의 건조 분말 제품(M-6) 265g을 수득하였다.This acid earthworm was then dissolved in a mixed acid of malic acid and lactic acid 1: 1.

The soil was excreted at a temperature of 15 ° C. for 2.5 hours to excrete soil in the digestive tract. The earthworms were then lightly washed and the mixer was wet milled. The suspension of the earthworms obtained was put in a tray and frozen at -35 ° C for 24 hours. Next, freeze-drying for 7 hours under vacuum of 0.1 mmHg at a temperature of −35 ° C., then raising the shelf temperature on which the tray is loaded to 22 ° C., vacuum drying for 10 hours under vacuum of 0.1 mmHg, and then increasing the temperature of the shelf. 265 g of an earthworm's dry powder product (M-6) was obtained by raising to 42 ° C. and drying for 15 hours under a vacuum of 0.2 mm Hg, and finally raising the temperature of the shelf to 78 ° C. for 7 hours under a vacuum of 0.1 mm Hg.

Example 23

에 인산 이수소칼륨 1. 5흘 용해하고 이것에 물로 가볍게 세정한 산지렁이(굵은 지렁이) 1kg을 넣고 10℃에서 2시간 방치하여 소화관 내의 분토를 배설시켰다.Acidic aqueous solution of pH 6.0 dissolved in citric acid 2. 5

Potassium dihydrogen phosphate was dissolved in 1. 5 days, and 1 kg of a mountain worm (coarse worm) lightly washed with water was added thereto and left at 10 ° C. for 2 hours to excrete the soil in the digestive tract.

Next, the mountain earthworms were washed twice with water, and the soils such as mud, powder and straw attached to the body surface of the mountain earthworms were washed out. Then it was wet milled with a mixer. Thereafter, 280 g of a dry powder of an earthworm was obtained by freeze vacuum drying the obtained suspension of the earthworm in the same manner as described in Example 21.

Example 24

(이중에 초산나트륨 1g과 황산나트륨 0. 59을 용존한다)중에 13℃에서 2. 5시간 방치하여 소화관 내의 분토를 배설시켰다.Dirt, powder, straw, etc. attached to the surface of 1 kg of earthworm (fishing earthworm) were washed well with water five times and washed. Next, the acid earthworm was dissolved in succinic acid at pH 5. 7 in acidic aqueous solution 3

The soil in the digestive tract was excreted by standing at 13 ° C. for 2.5 hours in 1 g of sodium acetate and 0.95 sodium sulfate.

Next, the mountain earthworm was washed with water, and then wet crushed with a homogenizer. Thereafter, 275 g of the earthworm dry powder was obtained by freezing and vacuum drying the suspension of the earthworm obtained in the same manner as described in Example 22.

Example 25

Earthworms (coarse earthworms) 1kg of dirt, dirt, straw, etc. attached to the surface of the body was washed well with water five times and washed. This acid earthworm was then dissolved in a mixed acid of citric acid and tartaric acid 1: 1. The soil in the digestive tract was excreted by standing at 15 degreeC for 2 hours in the acidic aqueous solution of 2.5 (9.g, which dissolved 0.7g of potassium citrate). The earthworms were then washed lightly with water and then wet ground with a blender. Thereafter, 283 g of earthworm dry powder was obtained by freeze vacuum drying the suspension of the earthworm obtained in the same manner as described in Example 22.

As described above, the present invention relates to a diabetes therapeutic agent comprising the dry powder of the earthworm or the extract of the earthworm. By administering the dry powder of the earthworm or the extract of the earthworm to experimental diabetic mice with alloxane, it was possible to significantly lower blood glucose levels.

Next, the volunteers of five diabetic patients were given an earthworm dry powder capsule (150 mg content) once a day, three times a day, orally and for 4 to 9 months with meals. 6 body bleeds after breakfast, 2 hours after lunch, 2 hours after lunch, 2 hours after dinner, 2 hours after dinner, every 1 month, 2 months or at least 1 month after 3-4 months And blood equivalents were measured. Sir this result. In patients with moderate diabetes mellitus, earthworms improved from 2 to 3 months after the administration of the earthworm dry powder, and from 4 months after the administration, the blood equivalents of all 6 items were standard values (standard values of blood sugar before breakfast, lunch, and dinner 3 meals were 50-100 mg ldl, The standard blood glucose level after 2 hours after each meal was lowered to within the range of 150 mg ldl or less). In severe diabetic patients, it was slightly difficult to drop the blood sugar levels of all 6 items to the standard level, but after 8 months of administration, the blood sugar levels of 4 items of 6 items were lowered to the standard value. In the long-term administration of earthworm dry powder for 6 to 9 months, there was no risk of hypoglycemia in which blood sugar dropped below the lower limit of the standard value. That is, by animal experiments and oral administration to humans, the dry powder of the earthworm of the present invention or the extract of the earthworm is safe and excellent diabetes treatment. I knew it was a preventive agent.

Claims (13)

An antidiabetic agent comprising the earthworm dry powder as an active ingredient. Mountain earthworms are left in an aqueous solution containing not less than 0.3% by weight of at least one compound selected from the group consisting of organic acid, inorganic acid, organic acid sodium salt, inorganic acid sodium salt, organic acid potassium salt and inorganic acid potassium salt or in fresh water. After excretion of the soil in the digestive tract of the earthworm, the soil attached to the surface of the mountain earthworms are removed by washing and removing the earthworm as it is, or the soil earthworms after washing and removing dirt attached to the body surface of the earthworm After excreting the soil in the digestive tract of the mountain earthworm by leaving it in aqueous solution or fresh water, the soil obtained by crushing the earthworm removed by washing and removing dirt from the body surface of the mountain earthworm with water was frozen at -10 to -60 ℃. Thereafter, freezing for 10 to 100 hours with a vacuum of 10 mmHg or less while gradually raising the temperature within the range of -16 to 80 ℃ The antidiabetic agent containing earthworm dry powder as an active ingredient characterized by carrying out the vacuum drying and drying the vacuum of the final process in a vacuum at 70 to 80 ° C. under a vacuum of 0.01 to 0.5 mmHg for 5 to 10 hours. . According to claim 2, the earthworms are left in the aqueous solution or in fresh water to excrete the soil in the digestive tract of the mountain earthworms, and the dirt adhering to the body surface of the mountain earthworms is washed and removed with water, and the grounded earthworms are crushed as they are. The diabetes therapeutic agent which performs the said process. The anti-diabetic agent according to claim 3, wherein the mountain earthworm before being left in the aqueous solution or fresh water is a mountain earthworm in which a part of the dirt is washed with water. The earthworm according to claim 2, wherein the soil adhering to the body surface of the mountain earthworm is washed off with water, and the earthworm is left in the aqueous solution or in fresh water to excrete the earthworm in the digestive tract of the mountain earthworm, and the ground earthworm is crushed as it is. Diabetes treatment agent to perform process. 6. The earthworm of claim 5, wherein the soil adhering to the body surface of the mountain earthworm is washed off with water, and the earthworm is left in the aqueous solution or in fresh water to excrete the soil in the digestive tract of the mountain earthworm, and washed with water. Cure as it is and diabetes treatment for performing the subsequent steps. The antidiabetic agent according to claim 2, 3, 4, 5 or 6, wherein the organic acid is acetic acid, citric acid, succinic acid, malic acid, tartaric acid or lactic acid. The agent for treating diabetes according to claim 2, 3, 4, 5 or 6, wherein the inorganic acid is phosphoric acid, sulfuric acid or hydrochloric acid. An anti-diabetic agent comprising as an active ingredient an extract obtained by extracting the earthworm with at least one extractant selected from the group consisting of an aqueous solvent, a water miscible organic solvent and a water immiscible organic solvent. The method of claim 9, wherein the aqueous solvent is fresh water, distilled water, physiological saline, buffer solution of pH 5 to 10 or prepared water of pH 5 to 10. The antidiabetic agent according to claim 9, wherein the water miscible organic solvent is an alcohol, ketone, ether or ester having 1 to 4 carbon atoms. The antidiabetic agent according to claim 9, wherein the water-immiscible organic solvent is a hydrocarbon having 6 to 10 carbon atoms, an alcohol, a ketone, an ester or a halogenated hydrocarbon having 1 to 5 carbon atoms. Mountain worms are left in an aqueous solution containing not less than 0.3% by weight of at least one compound selected from the group consisting of organic acid, inorganic acid, organic acid sodium salt, inorganic acid sodium salt, organic acid potassium salt and inorganic acid potassium salt or in fresh water. Earthworms in the digestive tract of the mountain earthworms are excreted, and the earthworm powders obtained by washing and removing dirts attached to the body surface of the mountain earthworms before and after this step with water, and then crushed and freeze-dried and vacuum dried are effective. A therapeutic agent for diabetes containing as an ingredient.