JPH02193929A - Diabete remedy - Google Patents

Abstract

PURPOSE:To provide a diabete remedy having a blood saccharide-reducing activity and safety by containing as an active ingredient an earthworm extract prepared by extracting an earthworm with an extraction solvent such as an aqueous solvent or a water-miscible organic solvent. CONSTITUTION:A diabete remedy contains as an active ingredient an earthworm extract prepared by extracting an earthworm with an extraction solvent such as an aqueous solvent, a water-miscible organic solvent or a water-non-miscible organic solvent. The earthworm is used e.g. in a form of an earthworm suspension prepared by leaving a living earthworm in an aqueous solution containing <=0.3wt.% of a compound such as an organic acid (e.g. acetic acid or citric acid), inorganic acid, organic acid sodium salt, inorganic acid sodium salt, organic acid potassium salt or inorganic acid potassium salt or in pure water to allow the earthworm to stew excrement soils in the digestive tracts, washing filth adhered to the surface of the body with water and subsequently wet- grinding the washed body.

Description

DETAILED DESCRIPTION OF THE INVENTION [Industrial Field of Application] The present invention relates to a therapeutic agent for diabetes. To be more specific, a diabetes treatment containing as an active ingredient an earthworm extract or a mixture of earthworm extract and earthworm dry powder, which has excellent blood sugar level (hereinafter referred to as blood sugar) lowering activity and safety. Regarding preventive agents.

[Conventional technology]

Dried earthworms have been used as medicine since time immemorial, mainly in Eastern countries, where they are referred to as kango and jiryu. The pharmacology and medicinal effects of earthworms reported in conventional literature are as follows.

■ “Worms and Life” by Mayu Obuchi, 1947 (Showa 22)
10330, published by Maki Shobo, pages 223-226, and “Fukukoku Mizu” by Shinkiji Hatai, April 30, 1980.
Published by Scientist Co., Ltd., pp. 160-163, earthworms are known to be effective in reducing bladder stones and expelling them from the body, as a treatment for smallpox, as a birthing agent, as a tonic, as a hair herbal medicine, as an aphrodisiac, and as an antipyretic. On the other hand, as earthworm toxins, it has been reported that one affects the nervous system and the other has a hemolytic effect, which is the destruction of red blood cells.

■ "The Dictionary of the People's Republic of China" edited by the Dictionary Committee of the Ministry of Health of the People's Republic of China, 1977 edition, some pages 197-19B states the following. Traditionally, earth dragon products have 2
There are different types. One such wide dragon (Lumbr icus)
Kwangtungasis) is prepared by tearing the abdomen open, washing away the internal organs and quicksand, and drying it in the sun, shade, or at low temperatures. Other Land Dragons (Lumbricus Nativ)
Earthworms are killed by placing them in plant ashes, then removing the ashes and drying them in the sun, shade, or at low temperatures, and their bodies are filled with mud. These two types of earth dragons are antipyretic,
4. As a convulsion treatment agent, blood circulation promoter, hemiplegia treatment agent, joint analgesic, urinary agent, bronchial asthma agent, and hypertension agent.
They report using 5 to 9 g/day.

■ “Our Chinese Herbal Medicine Series 3, Earth Dragon/Squid Bone/
Chinese Scientific Research, October 30, 1977, published by Matsuura Pharmaceutical Co., Ltd., page 7, reports that earth dragon tincture (ethyl alcohol extract of earth dragon) has a hypotensive effect.

■ “Natural Medicine Encyclopedia” edited by Takuo Okuda, April 15, 1986
Published by Hiroyo Shoten, p. 215, it is reported that earth dragon is used as a hypopyretic, analgesic, diuretic, and antidote.

■ Mamoru Tanaka [Hokkaido Medical Journal No. 24, pp. 18-24 (1949)] extracted kansei (dried fragments with mud removed) with boiled water and made a concentrate of this extract. The precipitated material obtained by adding ethyl alcohol to
It has been reported that when febrin) was dissolved in Ringer's solution and this solution was injected intravenously into an anesthetized cat, a rapid drop in blood pressure occurred, and an acceleration of blood coagulation was observed in proportion to the shock.

■ Kenji Iyou [Yamaguchi Igaku Vol. 9, pp. 571-576 (1960)] dissolved a physiological saline extract of the earth dragon, an ethyl alcohol or acetone extract of the earth dragon, and dissolved it in physiological saline. The solution was intravenously injected into adult rabbits and a drop in blood pressure was observed.

■ Volume 2 of "Middle and Weak Dictionary," edited by Jiangsu New Medical Academy, 1980, Published by Todo Science and Technology Publishing House, page 2112, includes Guangjilong tincture, suspension of dry powder of lily, and hot water soaking of lily. When administered to anesthetized dogs, large rats, cats, or mice with chronic renal hypertension, a slow and sustained blood pressure lowering effect was observed.

When earth dragon extract was injected intravenously into a dog or cat under anesthesia,
The blood pressure lowering effect appeared rapidly. However, it has been reported that it was not effective when administered orally or when applied clinically.

Furthermore, on page 2114 of the same magazine, it is stated that 40% concentration of Jiryu tincture (40g of Jiryu soaked in 60 degree ethyl chlorine loOm (2)) should be applied for 10 mQ each time, 3 times a day [i.e., Jiryu 1
This corresponds to 2g/day. (converted by the inventor)].

Those who cannot take tinctures can make pills by adding water to pure earth dragon powder (adding a small amount of excipients) and taking 3-4g each time.
- If taken three times a day [equivalent to 9 to 12 g/day (calculated by the inventor)] and continued for 30 to 60 days, it is effective against essential hypertension.

In addition, hypoxanthine was removed using Jiryu B1 liquid (HgC1), and the blood pressure lowering component was separated using an ion exchange resin.
If you take 1 mQ (containing 8 g of the herbal drug Jiryu) three times a day (equivalent to 24 g of Jiryu 7 days (converted by the inventor)), it is effective for essential hypertension. It is reported that

Conventional and customary methods for producing dried earthworm products or dry powder can be broadly classified as follows.

i. Rip open the abdomen of the earthworm and extract the internal contents (viscera and mud)
A method of removing and drying in the sun, shade, or at low temperatures (usually below 50°C).

i. A method of killing earthworms by placing them in plant ash, removing the ash and drying them in the sun, shade, or at low temperatures (usually below 50°C) to obtain earthworms with mud still stuck inside their bodies.

iii. After removing the mud inside the earthworm's body, the earthworm is placed in plant ash or fire ash and dried.

etc. These dried products were pulverized and used when needed or used. These manufacturing methods are simple and economical, and have the advantage of being easily carried out at home.

However, dried earthworm products or dried powders obtained by these methods can be stored in a refrigerator at 0 to 5 degrees Celsius or in an open state at room temperature of 5 to 45 degrees Celsius for up to about 6 months when stored in a closed condition. There is a drawback that mold may develop within a short period of one year or less, rendering the product unusable.

When earthworms are dried with their bodies filled with mud, as in method d above, or dried in plant ash or fire ashes, as in the melon manufacturing method, in most cases they are extracted with hot water when used as medicine. Alternatively, it is often decocted with boiled water, filtered, and the filtrate taken. In particular, the dried earthworm product or dry powder produced by the manufacturing method of H can be left as Jiryu tincture or powder.
Or it is rarely taken as a pill. In addition to taking the earthworms prepared by method 1 as a hot water immersion solution or a decoction, they can also be taken in the form of Jiryu tincture or Jiryu powder, or as a pill by adding a small amount of water or a small amount of excipient to the powder. There are many. When using the production methods i and i, the yield of dried earthworms with a moisture content of 10-16% from fresh earthworms is 5-9.
%, and the yield when using the same manufacturing method as H is 13 to 19%.

Recently, one of the inventors of the present invention, Yo Ishii, has developed a health food containing earthworm proteins and lipids as main components and a method for producing the same [Japanese Patent Application Publication No. 1983-216572 (publication date 1).
December 6, 984)].

This production method is an excellent method for obtaining dried earthworm powder as a health food. However, as a method for producing dried earthworm powder for use as drugs such as arteriosclerotic agents (also known as antihyperlipidemic agents), antidiabetic agents, and antihypertensive agents,
It was found that the medicinal efficacy was not sufficient. That is, when an external action is applied to remove excrement such as feces remaining in the body of a live earthworm, only the feces cannot be selectively removed. No matter how careful you are,
It has been found that the internal organs and body fluids, which contain many components that play an important role in medicinal efficacy, are removed along with the fecal soil, resulting in insufficient medicinal efficacy. Also, the yield for live earthworms is 10-1
Only 9%. Furthermore, a major problem is that the vacuum drying in the final finishing process is carried out for a long time of 20 hours or more at a vacuum level of 80°0. It was found that some enzymes were destroyed or inactivated.

Moreover, some of the other active ingredients were found to be thermally decomposed. Therefore, compared to the dried earthworm powder obtained by the production method of the present invention, the
The efficacy of the dried earthworm powder antidiabetic agent obtained in No. 16572 was approximately 50%.

Recently, Tsune Mihara, one of the inventors of the present invention, and other co-researchers have discovered that the fibrinolytic active substance of earthworms has an optimal pH of 8 to 1.
0. Stable pH is 5-1O, inhibited by Trasylol (trade name), Transamine (trade name), soybean trizcin inhibitor and serum, has plasminogen activation and fibrinolytic action, but has no fibrinogen dissolving action It was confirmed that it is an enzyme protein possessing various properties.

A patent application for a method for producing a fibrinolytic active substance consisting of a crude enzyme protein fraction extracted from earthworms using an aqueous solvent and a purified enzyme protein fraction obtained by purification thereof, ie, Japanese Patent Application Publication No. 148824/1983 (applied) February 27, 1982), U.S. Patent Application No. 470394 (filed on February 28, 1983) and British Patent Application No. 830359 (filed on February 25, 1983) claiming priority thereto. ), Italian Patent Application No. 4
7795A (filing date February 25, 1983), for French patent [No. 03165 (filing date February 2, 1983)
5th), West German Patent Application No. P3306944.1
(filing date February 28, 1983) and Canadian patent application N.
o, 422034 (filing date: February 21, 1983).

Furthermore, Tsune Mihara et al. applied for a substance patent for six new types of proteases from earthworms, namely, Japanese Patent Application Publication No. 1984-63184 (filing date, 1982).
(October 2, 2013) and a patent application for a thrombolytic agent containing these proteases as active ingredients, namely, Japanese Patent Application Publication No. 184131 (filing date: 1983).
March 31, 2013) and a foreign patent application claiming priority of these combined applications, namely Korean Patent Application No. 2990 (filing date 1
June 30, 1983) and the following patent applications were reported (A
U, P, App, l 6293 (filing date June 27, 1983), CA, P, App, 431387 (filing date 1
June 28, 983), DK, P.

App, 3008 (filing date June 29, 1983), E
P, P.

App, 83106288.0 (filed June 28, 1983), E S, P, App, 523754 (filed June 30a, 1983), F I , P, App, 83
2383 (filing date June 29, 1983), NO.P.
App, 2399 (filing date June 30, 1983), P
H, P, App, 29151 (filing date June 30, 1983), TW, P, App, 7211983 (
Application date: June 18, 1983), U.S., P., Ap.
p. 508163 (filed June 27, 1983)].

According to the research results of the present inventors, we found documents reporting the activity of a substance containing earthworm extract alone or a mixture of earthworm extract and dried earthworm powder as an agent for treating and preventing diabetes or as a hypoglycemic agent as an active ingredient. I couldn't. That is, we could not find any literature reporting that when administering a single earthworm extract or a mixture of earthworm extract and earthworm dry powder to experimentally diabetic mice induced by alloxan, the blood sugar level was significantly lowered.

Additionally, dried earthworm powder was administered to human diabetic patients for 4 to 9 months in addition to dietary therapy. In particular, in patients with mild to moderate diabetes, blood sugar levels improved after 2 to 3 months of administration, and after 4 months of administration, blood sugar levels were able to drop to normal levels for healthy people. We could not find any literature reporting excellent effects.

Furthermore, we could not find any literature reporting a method for producing earthworm dry powder and earthworm extract having such effects.

[Problem that the invention seeks to solve]

Traditionally, oral antidiabetic agents include sulfonylurea and biguanide (
Organic synthetic compounds such as biguanide compounds are widely used.

The present inventors have been intensively researching the use of dried earthworm powder for many years with the aim of creating highly safe medical products free of side effects from natural products rather than from such synthetic organic compounds. It was surprisingly discovered that an earthworm extract or a mixture of an earthworm extract and a dried earthworm powder is effective for the treatment and prevention of diabetes, and the present invention was completed.

[Failure to solve the problem]

A detailed study was conducted to prepare a single earthworm extract or a mixture of earthworm extract and earthworm dry powder that is safe and has excellent antidiabetic effects.

The earthworm extract of the antidiabetic agent of the present invention is prepared by using earthworms as a raw material in an aqueous solvent, a water-miscible organic solvent (meaning an organic solvent that is miscible with water), and a water-immiscible organic solvent (meaning an organic solvent that is miscible with water). It is an extract obtained by extraction treatment with at least one type of extractant selected from the group consisting of: This earthworm extract can be obtained by the following production method.

That is, raw earthworms are extracted with at least one type of 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 form a precipitate. Alternatively, an organic solvent was added to the concentrated extract to obtain a precipitate, or an extract was obtained by completely removing the extraction solvent under normal pressure, increased pressure, or reduced pressure.

The earthworms used as raw materials are dried earthworms and dried powder thereof obtained by the known manufacturing method of I and Yama of the previous period and the manufacturing method described in Japanese Patent Application Publication No. 59-216572; live earthworms from which excrement has been removed; excrement from which they have been removed. Shitanama worms (meaning earthworms whose excrement is removed from the earthworms as they are collected, and which are not boiled, baked, dried, or pickled.); Live earthworms from which excrement has been removed, or the form of a crushed suspension of raw earthworms. earthworms can be used.

As a result of detailed research, the present inventors have found that in order to obtain an earthworm extract that is completely sterile, has excellent hypoglycemic activity, and does not have the odor inherent to earthworms, it is necessary to extract raw earthworms ( It has been found that a combination of a purification method or cleaning method for harvested or cultured earthworms, etc., and freezing and vacuum drying conditions is a particularly important operation. According to actual measurements by the present inventors, the filth attached to the body surface of raw earthworms, such as collected or cultured earthworms, is generally
40% (by weight), and the excrement in the fire extinguishing pipe contains 5 to 15% (by weight) based on the wet earthworms. We have discovered that using clean earthworms from which these filth and excreta have been promptly removed as raw materials for extraction is an important factor in improving medicinal efficacy and yield. That is, the most preferable raw material earthworms are earthworm suspensions obtained by wet pulverization in the processing steps of production methods 1 and 2 described below, and dry earthworm powders produced by freezing and vacuum drying these suspensions. As shown below, the earthworm extract prepared from the earthworm suspension and earthworm dry powder obtained by the methods 1 and 2 has a blood sugar-lowering activity superior to that of the earthworm dry powder. Furthermore, it was found that a mixture of earthworm extract and earthworm dry powder had even better hypoglycemic activity than earthworm extract alone.

Below, the method for preparing the starting materials, which is an important operational step in the manufacturing method of the product of the present invention, will be explained.

Method for producing earthworm suspension and dry earthworm powder - Method 1: Live earthworms are mixed with acetic acid, citric acid, conolic acid, malic acid,
Non-toxic organic acids such as tartaric acid, lactic acid or phosphoric acid, sulfuric acid,
At least one compound of non-toxic inorganic acids such as hydrochloric acid or sodium or potassium salts of these acids at 0.3
(weight)% or less, preferably 0.1 (weight)% or less, in a slightly acidic aqueous solution or fresh water with a pH of 3 to 6.5 at a temperature of 1 to 25°C for 0.5 to 48 hours, preferably is left at a temperature of 8 to 20°C for 1 to 24 hours to fully excrete the fecal soil in the digestive tract of the live earthworm using its own excretory power, and then it is attached to the body surface of the live earthworm with water. The filth was washed off, and then wet pulverization was performed to obtain a suspension of earthworms. This earthworm suspension was frozen at a low temperature of -5°C or lower, preferably -60°C, and then subjected to freezing and vacuum drying. The conditions for freezing and vacuum drying are temperature -60 to +90℃, vacuum 100nI!
1Hg or less, preferably 30mmH at -40-+80℃
5 to 100 g while increasing the temperature stepwise at a vacuum level of less than 5 g.
Freezing and vacuum drying were carried out for preferably 10 to 60 hours to obtain a sterile dry powder of earthworms.

Manufacturing method 2: After washing off the dirt adhering to the body surface of live earthworms with water, use non-toxic organic acids such as acetic acid, citric acid, succinic acid, malic acid, tartaric acid, lactic acid, or phosphoric acid, sulfuric acid, hydrochloric acid, etc. at a low concentration of 0.3% (by weight) or less, preferably 0.1% (by weight) or less, or at a pH of 3 to 6. The living earthworms are allowed to stand in a slightly acidic aqueous solution or in fresh water at a temperature of 1 to 25°C for 0.5 to 48 hours, preferably at a temperature of 8 to 20°C for 1 to 24 hours, using the excretory power of the living earthworms themselves. After sufficiently excreting the fecal soil in the digestive tract of the live earthworms, wet crushing was performed to obtain an earthworm suspension. This earthworm suspension was frozen at a low temperature of -5°C or lower, preferably -10 to -60°C, and then subjected to freezing and vacuum drying. The conditions for this freezing/vacuum drying are a temperature of -60-+90°C, a vacuum of 100 mmHg or less, preferably -40 to +80'0, and a vacuum of 30 mmHg or less for 5 to 100 hours, preferably 10 Freezing and vacuum drying were performed for ~60 hours to obtain a sterile dry earthworm powder.

The wet grinding method of earthworms, that is, the method of destroying earthworm tissues (cells), can be made into a suspension or homogeneous liquid using equipment such as a homogenizer blender, homomixer, crusher, or pressurized cell destruction device. preferable. The temperature during this wet grinding is preferably 1 to 25°C, preferably 2 to 15°C.

By the operation in the first stage, it was possible to obtain yellowish brown or brown earthworm dry powder from living earthworms at a yield of 20 to 35%.

Normally, the moisture content of this dry earthworm powder is 5 to 16%, preferably 7 to 14%, and the ash content is 3 to 8%, preferably 4 to 7%.
%, nitrogen content was adjusted to 1 to 11%, preferably 6 to 11%. In addition, the dried earthworm powder contains aspartic acid, threonine, serine, glutamic acid, proline, glycine, alanine, cysteine, valine, methionine,
Contains 18 or more amino acids: inleucine, leucine, tyrosine, phenylalanine, tryptophan, lysine, histidine, and arginine.

Test Example 1 Table 1 shows the crude analysis results of the dried earthworm powder obtained by the methods of the first half production methods 1 and 2 and the production method of the examples described later.

Test Example 2 The results of component analysis of dry earthworm powder products M-2 and M-4 obtained by the method of Production Method 1 and M-5, M-6, M-8 and M-9 obtained by the method of Production Method 2 were It is shown in Table-2.

(Leaving space below) Test Example 3 Amino acid analysis of the crude protein compartment in the dry earthworm powder products of M-2 and M-4 obtained by the method of Production Method 1 and M-6 obtained by the method of Production Method 2 was conducted. Table 3 shows the results of comparison with the analytical values of fishmeal and soybean flour.

(Left below) Table 3 *P, Mc Donald, R, A, Edward
s and J, F, D Greenhalgh: A
From data described in normalNutrition (1973).

From Tables 2 and 3, it was found that the dried earthworm powder obtained by Production Methods 1 and 2 contained abundant crude protein, crude lipids, and various metals. The amino acid composition revealed that it contains a large amount of essential amino acids.

Production method 1 yielded a slightly more preferable product than the dried earthworm powder obtained using production method 2. Furthermore, it is better to leave live earthworms in an aqueous solution containing at least one compound of the above-mentioned organic acid, inorganic acid, or sodium or potassium salt of both of these acids at a low concentration as above, rather than leaving them in fresh water. The excretion of fecal soil in the digestive tract of live earthworms is faster and the excretion rate is higher than that of live earthworms. According to research conducted by the present inventors, when live earthworms are left in fresh water at a temperature of 8 to 18 degrees Celsius, it takes 15 to
It took 48 hours. In this regard, it has been found that when left in the above-mentioned dilute aqueous solution at 8 to 18°C, the excrement is completely discharged in 0.5 to 5 hours.

In the above-mentioned production methods 1 and 2, the finished state of the dried earthworm product after drying is often in an aggregated state or a state in which clumps are mixed. However, when this is applied to a pulverizer, it is easily turned into powder. When administering dried earthworms and earthworm extracts to mammals including humans, powders are preferable to lumps.

Tsune Mihara et al. [Japanese Patent Application Publications Nos. 59-63184 and 59-184131] fractionated six types of new proteases, fibrinolytic enzymes, from earthworms. Namely, Tsune Mihara et al. added 10 times the amount of physiological saline to dried earthworm powder and incubated it for 2 days. After carrying out ammonium sulfate fractionation on the supernatant fluid, the precipitate was fractionated with 5ephacryl S-200. Gel filtration was performed, and DEAE-cellulose ion exchange chromatography was performed on the resulting protein fraction, resulting in proteins in fractions 11 (1) and (2) having casein-degrading and fibrin-degrading activities. Further DE regarding this r, n, m fraction
AE cellulose, 5ephadex G-75, Toyovar HW55, A CH-SepharoseSB
As a result of purification using enzamidine-Sepharosa or the like, 6 fractions of purified enzyme were obtained. 5
When the molecular weight was measured by DS-PAGE, the molecular weight of fraction 1-0 was the lowest, calculated as 23.500, and then the molecular weight was sequentially increased to I-l5I-2, I[, ■-1, m-2. The molecular weight of lll-2 was 34.200. Also, when measuring the isoelectric points of these 6 fractions by isoelectric focusing, I
-0 is the highest, pH 4°12, then successively p
H was low, pH 3.52 at m-2. These 6 fractions are new proteolytic enzymes that are different from serine enzymes, and the optimal pH of the proteases in these 6 fractions is
It is reported that the stable pH was 4-12 or 5-12, the optimum temperature was 50°C or 50-60°0, and the inactivation conditions were 70°C for 60 minutes.

Dry earthworm powder product M-4 obtained by the above production methods 1 and 2
When 10 times the amount of physiological saline was added to M-6 and the supernatant was measured using a standard fibrin plate, fibrinolytic activity was immediately observed as shown in Table 4. When this physiological saline solution of M-4 earthworm dry powder is incubated at 37°C, the fibrinolytic activity of the supernatant increases by about 4 times on the 10th day, as shown in Table 4, and on the 50th day. 5 times on the 75th day, and 5.5 times on the 75th day.

This fact suggests that a large amount of the precursor of this protease exists in the freeze-dried earthworm powder, and the enzyme activity is expressed through autolysis.

When the activity of the supernatant on day 50 was converted into international units of urokinase and compared, it was calculated to be approximately 8.0001010+12. In addition, this enzyme dissolves both plasminogen-free fibrin plates and standard fibrin plates, and the standard fibrin plate has a larger dissolution window than the glasminogen-free plate, and has enzyme activity that directly degrades fibrin, as well as plasminogen activator activity. was also shown. In the above operation, the same results as shown in Table 4 were obtained when M-6 earthworm dry powder was used instead of M-4.

(Margins below) Table 4: Add 10 times the amount of physiological saline to the dry earthworm powder,
For details on the reason why fibrinolytic active earthworm dry powder and earthworm extract of the supernatant after incubation at 37°C exhibit diabetes therapeutic/preventive effects or hypoglycemic effects when orally administered to rats and humans, Although it is unknown,
The proteolytic enzyme (protein) itself, the precursor of this enzyme (protein) itself, or the protein, glycoprotein, metal-binding protein, lipid, carbohydrate, or other unknown compound itself contained in the dried earthworm powder and earthworm extract. Or, the nitrogen content of the most preferred earthworm extract and earthworm dry powder as a therapeutic agent is 7 to 16%, that is, the crude protein content is 43%.
．． 8-100%.

Next, preferred specific examples of freezing and vacuum drying operations of the suspension obtained by wet pulverization of earthworms obtained by the above-mentioned production methods 1 and 2 are as follows.

Wet ground earthworm material, that is, earthworm suspension, is
After freezing at -60°C, preferably -30°C to -50°C for 5 to 60 hours, the temperature is 0. Lyophilize for 5-12 hours under vacuum at O1-0.2 mmHg. Next 20-30
Dry under vacuum at 0.01-0.2 mmHH for 5-15 hours at <0>C. Next, 35-50℃, 0.1-0.5mmH
Dry under g vacuum for 10-20 hours. Vacuum drying in the next final finishing step is carried out under a vacuum of 0.01 to 0.5 mmHg at 70 to 80°C, preferably at 75 to 80°C for 5 to 10 hours. A dry powder could be obtained. In particular, the final finishing step, vacuum drying, is also an important operation. In order to make the proteolytic enzymes, enzyme precursors, and other dry powders contained in dried earthworm powder aseptic, the present inventor conducted detailed research and found out the vacuum drying conditions for the final step of freezing and vacuum drying. It was discovered that the combination of the three elements of vacuum degree, heating temperature, and time was an important condition, and the above operating conditions were established.

As shown above, six types of purified proteolytic enzymes from dried earthworm powder (Japanese Patent Application Publication No. 59-6318
It has been reported that the proteolytic enzymes in the dried earthworm powders obtained by Production Methods 1 and 2 are deactivated in 60 minutes at 70°C, as shown in Table 4. Not being utilized. In addition, the dried earthworm powder obtained by Production Methods 1 and 2 was kept at room temperature of 5 to 45°C in a sealed state for 5 years.
In the preserved examples, no mold growth or other deterioration was observed.

The most preferred raw materials in the present invention are a suspension of earthworms obtained by wet pulverization in the processing steps of Production Methods 1 and 2 as described above, and a dried powder of earthworms produced by freezing and vacuum drying the suspension. . The next preferred raw material earthworms are the above-mentioned publicly known earthworms and the Japanese patent application publication No. 5
These are dried earthworms and their dried powder obtained by the manufacturing method of No. 9-216572. However, the latter three types of raw earthworm extracts are considerably inferior in medicinal efficacy to the former two most preferred raw material extracts, about 30 to 50%
Only 0%. The reason is as follows.

That is, when an external action is applied to remove the manure remaining in the digestive tract of the earthworm (containing about 5 to 15% based on wet earthworms), only the manure cannot be selectively removed. No matter how carefully they are handled, internal organs, digestive juices (including various effective enzymes such as cellulases, amylases, proteases, and lipases) and body cavity fluids that contain many proteins play an important role in medicinal efficacy. (
Coelonic Nuid) and other body fluids are removed from the body along with the fecal soil, resulting in insufficient medicinal efficacy. In addition, as shown in the process description and examples of manufacturing methods 1 and 2 above,
When raw earthworms are left in a dilute aqueous solution, it is important for medicinal efficacy that the feces in the earthworm's digestive tract be completely expelled within the shortest possible time. When the dilute aqueous solution is a concentrated aqueous solution, the earthworms that have been left unattended will have to remove the soil from the back of the earthworm's body (segment) before excreting the excrement.
Yellow or milky coelomic fluid (Coelonic fluid) from the dorsal foramen (Dorsa 1pore) in the dorsal foramen (sum)
The concentration of the dilute aqueous solution should be set to 0.
There is a reason to limit it to a low concentration of 3% (by weight) or less or a slightly acidic pH of 3 to 6.5.

When earthworms are left in a dilute aqueous solution or fresh water for a long time, for example, for 3 days (72 hrs), they excrete a large amount of body fluids such as digestive juices and body cavity fluids in addition to feces, resulting in a decrease in body weight. At this time, the weight of the earthworm decreases in inverse proportion to the length of time it is left in the water. Dry powders produced using such earthworms as raw materials and earthworm extracts thereof lack medicinal efficacy. Therefore, as shown in the examples of the present invention, earthworms that can sufficiently (completely) expel the excrement from the digestive tract within a short period of time are preferable. Further, dirt adhering to the body surface of earthworms can be relatively easily removed by washing with clean water. Among the two types of preferable raw material earthworms mentioned above, a particularly preferable raw material form is to leave live earthworms in the above dilute aqueous solution to dislodge the excrement in the digestive tract of the live earthworms, and then wash them with water and subject them to wet crushing. This is a paste-like suspension of earthworms obtained through this process.

When this suspension is used as a raw material, the amount of solvent used can be saved compared to other forms of earthworms, the active substances can be extracted within a short time, and the amount of extract per unit of processing time is large. It has excellent extraction efficiency.

The aqueous extraction solvent is fresh water, distilled water, physiological saline, p
It is preferable to use various buffer solutions of H5 to H10 or various prepared waters. The buffer solution is phosphoric acid, acetic acid, boric acid, citric acid, Tris/hydrochloric acid, etc. at pH 5 to IO, preferably pH 6 to
One or more of the 8 buffer solutions having various compositions can be used. Also, p
H5-1O Preferably, the pH value of the prepared solution is 6-8, which is a mixture of a water-soluble inorganic acid such as hydrochloric acid, sulfuric acid, phosphoric acid, acetic acid, lactic acid, citric acid, succinic acid, or tartaric acid, or an organic acid and an alkali metal such as sodium or potassium. Refers to dilute aqueous solutions prepared in-house from hydroxides or carbonates. The most preferred aqueous solvents are saline, buffer, and fresh water. When using these aqueous solvents, it is preferable to use a preservative such as parahydroxybenzoic acid ethyl ester, benzoic acid sodium salt, sodium azide, etc. in an amount of 01 to 0.3% (by weight).

Examples of water-miscible organic solvents include methyl, ethyl,
Allyl, n-propyl, i-propyl, n-butyl, S
-Butyl or L-butyl alcohol with 1 or more carbon atoms
Lower alcohols of 4; Ketones having 3 to 4 carbon atoms such as acetone or methyl ethyl ketone: ethyl ether, 1
．． It is preferable to use ethers having 3 to 4 carbon atoms, such as 2-propylene oxide, dioxolane, 4-methyldioxolane, or dioxane; and esters having 2 to 4 carbon atoms, such as methyl formate, ethyl formate, and methyl acetate.

Among these, the most preferable water-miscible organic solvent has 1 carbon number.
~4 alcohols.

Examples of water-immiscible organic solvents include n-hexane,
Hydrocarbons having 6 to 10 carbon atoms such as i-hexane, n-hebutane, i-hebutane, octane, i-octane, cyclohexane, benzene, toluene, xylene or ethylbenzene; n-hexyl, 2-methylpentyl, 2-
Alcohols with 6 to 10 carbon atoms such as ethylbutyl, S-hexyl, S-heptyl, n-octyl, 2-ethylhexyl, 2,6-cymethyl-4-heptyl, n-decyl alcohol; methyl-n-amylketone, m- or p
- Ketones having 7 to 8 carbon atoms such as methylcyclohexanone or methylhexyl ketone; n-butyl acetate, S acetate
-butyl, i-butyl acetate, 2-ethylbutyl acetate,
Esters having 6 to 10 carbon atoms such as S-amyl acetate, methylamyl acetate or 2-ethylhexyl acetate; halogenated hydrocarbons having 1 to 5 carbon atoms such as chloroform, dichloropentane or epichlorohydrin are preferably used. .

The extraction method may be carried out by a general method and is not particularly limited, but examples thereof are as follows.

For raw earthworms in the above form, an aqueous solvent, concentration 5 to 7
from the group consisting of an aqueous solution of an organic solvent at a concentration of 0% (by volume), preferably from 10 to 60% (by volume), a water-miscible organic solvent at a concentration of 80-100% (by volume), a water-immiscible organic solvent at a concentration of about 100%. At least one type of extractant selected from 1 to 100
twice the amount (volume/weight), preferably 2 to 30 times the amount, the extraction temperature is -40 to +60°C, the extraction contact time is from 10 minutes to about 100 days, preferably from 30 minutes to about 40 days,
After occasional or continuous stirring, the mixture was allowed to stand and filtered to separate the extract. For filtration, a pressurized or reduced pressure filter (machine) or a centrifugal separator may be used.

After the extraction residue is thoroughly washed, the extract and washing liquid are combined. An extract was obtained by removing the extraction solvent from the extract or combined solution at a temperature of 60° C. or below under normal pressure, increased pressure, or reduced pressure, by a concentration method such as ultrafiltration concentration, or freeze-drying. However, when ethanol, for example, is used as the extractant, depending on the type of final target side, the ethanol may not be completely removed and may remain. Alternatively, a crystal precipitation method may be used in which the concentrated solution of the extract or mixture is allowed to stand, or a method in which an organic solvent is added to the concentrated solution to precipitate crystals, followed by filtration and drying to obtain an extract.

The yield of the extract based on the raw earthworms in the above operation is 3 to 60% (by weight) in terms of dry matter. This difference in yield depends on the form of the raw earthworm, the type of solvent, extraction temperature, extraction time, and other extraction conditions. If the extraction temperature is high, there is a risk that the active substance will be decomposed, so the extraction temperature should be between -40 and +6.
0°C is preferred.

The components contained in the extract are as shown in Table 2 and Table 3.
It contains 18 types of amino acids and 8 types of metals, just like the ingredients in dry earthworm powder. In particular, the proteins in this extract include proteins made of the same amino acids as the raw earthworms, carbohydrate proteins, metal-binding proteins, and the like. Furthermore, the extract contains free amino acids, lipids, various carbohydrates, ATP and A
Contains TP-like substances, various enzymes, and other effective chemicals of unknown composition.

Production method 3 Preparation of raw earthworms and dry earthworm powder: Live earthworms (
The red earthworm) lokg was left in an acidic aqueous solution 2012 with a pH of 5.8 in which a mixed acid of 2:1 of malic acid and lactic acid was dissolved at a temperature of 10° C. for 2 hours to sufficiently excrete the feces in the digestive tract. Wash the live earthworms thoroughly with water to remove dirt, feces, and other dirt adhering to the body surface of the live earthworms.Ultra Homo Mixer (manufactured by Nippon Seiki Co., Ltd.)
Wet-grind with f-mi.

9.3 kg of a suspension of water was obtained. Add this earthworm suspension to 1.
Divide into 6 equal parts of 55 kg and divide each into E-1, -2, -
3, -4, -5 and -6.

1.55k of E-1 and E-2 earthworm suspension obtained above
Place g in each tray and freeze at -40°C for 30 hours.

The product was then freeze-dried at a temperature of -40°C under a vacuum of 1mmHg for 6 hours, and then at 25°C for 0.5 hours. After drying under a vacuum of lmmHg for 8 hours, then drying at 45°C for 12 hours under a vacuum of Q, lmmHg, and finally at 80"0 to 0.lmHg.
By drying under vacuum for 6 hours, 318 g of dried earthworms were obtained in each of the two trays. Let this be ni-1 and ni-2.

Preparation of extract:
After extracting 3 times at 5°c-c, the residue was finally diluted with 3 times the amount of 9
It was washed with 0.945Q of 9% methanol, and the combined liquid of the extract and washing liquid was dried under reduced pressure at 30°C, and then vacuum-dried to obtain 88.2 g of a powder extract. This is extract
■.

X-■, dried earthworms (K-2) 315gin, 3
A combined solution of twice the amount of chloroform 0.945m and twice the amount of ethanol 0.63Q was added and extracted three times at 20°0.Finally, the residue was washed with twice the amount of ethanol 0.63Q, and the extract The combined solution of the extract and the washing liquid was concentrated to dryness under reduced pressure at 30° C., and then vacuum-dried to obtain 72.5 g of a powder extract. This is referred to as extract X-■.

X-■, earthworm suspension (E-3) 1.55 kg+:
After adding 6.2Q of fresh water and stirring at 10℃ for 2 hours,
Filter and separate into extract and residue. After washing the residue with water, a combined solution 7.7Q of the extract and washing solution was obtained. This mixture 7.7
Q1. : After adding 2.31Q of n-hexane and stirring, leave to stand and separate into the water-soluble layer and the n-hexane layer, separate the water-soluble layer, concentrate to dryness under reduced pressure at 30°C, and then freeze-dry. 86.8 g of powder extract was obtained. This is extracted as Extract
■.

X-■, 4 to 1.55 kg of earthworm suspension (E-4)
Add 10 ff of 0% ethanol aqueous solution, stir at 40°C for 12 hours, filter, and separate into extract and residue.

The residue was washed with 0.3α of 40% ethanol, and the combined liquid of the extract and washing liquid was concentrated to dryness under reduced pressure at 40° C., and then vacuum-dried to obtain 69.7 g of a powder extract.

This is referred to as extract X-■.

X-■, To 1.55 g of earthworm suspension (E-5), 2.5 Q of pH 6.4 phosphate buffer was added, and the mixture was heated at 37°C.
After incubating for 8 hours to promote the action of protease naturally present in the earthworm's body, it was filtered, the residue was washed with a phosphate buffer solution of pH 6.4, and the extract and washing solution were combined into 3. Got 9Q. After adding sodium azide Ig to this mixture and incubating at 37°C for 8 hours, it was concentrated under reduced pressure at 30°C. Two volumes of ethanol were added to the resulting concentrated solution, the precipitate was filtered, and the precipitate was fractionated. was vacuum dried and then lyophilized to obtain 64.2 g of powdered extract.

This is referred to as extract X-■.

X-■, After naturally drying 1.55 kg of earthworm suspension solution (E-6) in the sun, powdered earthworm dry powder (
Named K-3. ) to ethanol and ethyl ether 2:1
A mixed solvent of 900°C was added and extracted three times at 20°C. Finally, the residue was washed with the mixed solvent from the previous period, and the combined liquid of the extract and washing liquid was concentrated to dryness under reduced pressure at 30°0, and then vacuum-dried to obtain 61.5 g of extract. This is extract
−■.

Manufacturing method 4 Raw earthworms and earthworm drying - Preparation of powder: Wash 13 kg of cultured live earthworms (red earthworms) with mud thoroughly with water to remove the mud adhering to the body surface, and then add malic acid and citric acid. pH 6.2 with 1:l mixed acid dissolved
The live earthworms were left in an acidic aqueous solution 40Q for 2 hours at a temperature of 12°C to fully excrete the upper part of the digestive tract, and then the live earthworms were thoroughly washed with water to remove the mud attached to the body surface of the live earthworms.
After washing off dirt such as feces, the live earthworms were wet-pulverized in a mixer to obtain 9.01 τg of mixer suspension.

This earthworm suspension was divided into 6 equal parts of 1.5 kg each, and E=7, -8, -9, -10. -11 and -12.

1.5 kg of the E-7 and E-8 earthworm suspensions obtained above were placed in each tray and frozen at -30°C for 40 hours.
Next, freeze-dry the product at a temperature of -40℃ for 6 hours under a vacuum of lmmHg, then increase the temperature of the shelf on which the tray is placed to 30℃.
After drying under a vacuum of lmmHg for 6 hours, the shelf temperature was raised to 50 °C and vacuum dried under a vacuum of 0.2 mmHg for 8 hours, and finally the shelf temperature was raised to 78 °C and dried under a vacuum of 0.2 mmHg for 8 hours. By drying for hours, 305 g of dried earthworms were obtained. Let this be -7 and -8.

Preparation of extract: X-■, 300 g of dried earthworm (K-7): 3
0.9% aqueous sodium chloride solution containing 0.02% parahydroxybenzoic acid ethyl ester was added thereto, stirred at 30°C for 96 hours, filtered, and the residue was added to the 0.02% parahydroxybenzoic acid ethyl ester solution.
The extract was washed with a 9% aqueous sodium chloride solution, and a clear extract 3.8Q was obtained by combining the extract and the washing liquid.

This was concentrated by ultrafiltration to a liquid volume of 0.16ff,
Ethanol O, 16Q was added to this and precipitated and fractionated, and the resulting precipitate was added to the filtrate so that the final concentration of ethanol was 80%, and the obtained precipitate was vacuum-dried to obtain a crystal powder of 14.4
Obtain gI;. This is designated as extract X-■-1.

The filtrate of extract X-■-1 was concentrated to dryness under reduced pressure at 30°C to obtain 72.5 g of crystalline powder. This is extract
-■-2.

X-■, 3Q of 0 to 300g of dried earthworms (K-8)
．． Add fresh water containing 1% sodium benzoate and heat at 32°C.
After stirring and extracting for 72 hours, the residue was filtered and the residue was reduced to 0.
The extract was washed with 6Q fresh water, and the extract and washing liquid were combined to obtain a clear extract 3.5α. This extract was concentrated by ultrafiltration and then concentrated to dryness under reduced pressure to obtain 69 g of crystalline powder. This is referred to as extract X-■.

X-■, 40 kg of earthworm suspension (E-9)
Distilled water containing 0.02% parahydroxybenzoic acid ethyl ester of 3Q was added to the dried earthworm powder (referred to as K9) that had been vacuum-dried at 32°C.
After stirring for a while, filter and separate the extract and residue. After thoroughly washing the residue with distilled water, the combined liquid of the extract and washing liquid was concentrated under reduced pressure at 30°C to about 1/10 the volume. After adding 4 times the volume of ethanol and stirring well, the mixture was allowed to stand overnight. The generated precipitate was filtered, washed with ethanol, and then vacuum dried to obtain 72.5 g of a powder extract. This is referred to as extract X-■.

X-■, earthworm suspension (E-10) 1.5 kg to 6
7.5Q of 0% ethanol aqueous solution was added, and the mixture was allowed to stand at 20 to 22°C for 8 days with occasional stirring in a sealed state.

After that, it was filtered and the residue was dissolved in 0.5% 50% ethanol aqueous solution.
12, and the combined liquid of the extract and the washing liquid was concentrated to dryness under reduced pressure at 30° C., and then freeze-dried to obtain 40.5 g of a powder extract. This is designated as extract x-■.

X-■, earthworm suspension (E 11) 5 to 1.5 kg
A mixed solvent of 4.5Q of 0% ethanol aqueous solution and 3Q of 50% Impropatool aqueous solution was added, and the mixture was left standing at 8° C. for 166 days with occasional stirring in a tightly stoppered state.

After that, it was filtered and the remaining liquid was dissolved in 50% ethanol aqueous solution (0.5%).
After washing with M, the combined liquid of the extract and washing liquid was concentrated to dryness under reduced pressure at 35°C, and then freeze-dried to obtain 28.2 g of a powder extract. This is referred to as extract X-■.

X-■, 1.5 kg of earthworm suspension (E-12)
Add 7.5Q of 0% ethanol aqueous solution and extract 3 times while stirring at 25°C for 2 hours.Finally, wash the residue with 60% ethanol aqueous solution 1 (2), and add 30% of the combined extract and washing liquid.
90°C by concentrating to dryness under reduced pressure and then freeze-drying.
．． 5g of powder extract was obtained. This is designated as extract x-■.

The earthworms used in the present invention are red earthworms (Lumb.
ricus rubellus), LT earthworm (also known as Lumbricus tsrrestris)
)), striped earthworm (Eiseniafoetida),
Ailolobophorac (AIlolobophorac)
at 1g1nosa), Purple Treeworm (Den
drobaenaoctaedra), cherry earthworm (
Allolobophora japo-nica M
ichhaelsen), ground worm (Drawida)
hatta-mimizu Hatai), Pheretima diver-gens M
ichalsen), common earthworm (Pheret)
imacommunissima), Hatagamagamizu (P
Heretima agres-tis), Siebold earthworm (Pheretima 5ieboldiHor)
st), Pheretima hi
lgen-dorfi), Isoearth earthworm (PonLodr
illus matsushimensisI 1zuk
a), Tubifex h
atLai Nomura), Limnodri lusgotoi (also known as Limnodri lusgotoi)
Hatoi=L, 5ocialis 5tephe
Any normally growing earthworm can be used.

When administering the earthworm extract or the mixture of earthworm extract and earthworm dry powder of the drug of the present invention for clinical treatment, the form may be either oral or parenteral, but oral administration is particularly preferred. The oral dosage forms of the present invention include capsules, tablets, granules, powders, coatings, sugar-coated tablets, and emulsions. Such formulations are used. Examples of pharmaceutical carriers include excipients such as lactose, sucrose, mannitol, glucose, starch, sorbitol, glycine, calcium phosphate, and microcrystalline cellulose; binders such as starch, gelatin, acacia, glucose, sucrose, sorbitol, and mannitol. , tragacanth, hydroxypropylcellulose, hydroxypropoxymethylcellulose, carboxymethylcellulose, 2-methyl-5-vinylpyridine-methacrylic acid-methylethyl acrylate copolymer, polyvinylpyrrolidone, sodium alginate, etc.; stearic acid as a lubricant, curing oil, magnesium stearate, calcium stearate, polyoxyethylene monostearate, talc, silicon oxide,
Polyethylene glycol, etc.; starch containing potato flour, surfactant, etc. as a disintegrant; sodium lauryl sulfate, etc. as a wetting agent. Furthermore, when administered parenterally, it can be used fully. In particular, cocoa butter, Witebsol (Wit) is used as a base for full expansion.
Epsol, Subanal, polyethylene glycol, polypropylene glycol, glycerogelatin, gelatin capsules, etc. are used. In addition, known safe preservatives such as methyl parahydroxybenzoate, propyl parahydroxybenzoate, butyl parahydroxybenzoate, butyl hydroxyanisole, and other safe dyes are used in combination.

The dosage of the earthworm extract alone or the mixture of earthworm extract and earthworm dry powder of the antidiabetic agent of the present invention varies depending on the administration method, the age, weight, condition, and type of disease of the patient, but is usually human. About 0.01g to 5g per day is preferable. Most preferably, the dose is 0.02g to 2g per day, divided into 1 to 3 doses per day.

Effects: The pharmacological test methods and results of the toxicity and antidiabetic effect or blood sugar lowering of the earthworm extract, the mixture of the earthworm extract and the dried earthworm powder, and the dried earthworm powder of the present invention will be described in detail below.

A. Acute toxicity test of dry earthworm powder DDY male base weighing 30 ± 2 g and body weight 100 ± 2
An acute toxicity test by oral administration was conducted using 5 mice in each group of Wistar base rats. Earthworm dry powder M-1 (contains 10.2% moisture, 5.1% ash, 9.4% nitrogen); FIM-2 (10.4% moisture, 5 ash)
．． 3%, nitrogen 8.6%); same M-3 (moisture 10.7
%, ash content 5.2%, nitrogen content 9.2%); Same M-4 (moisture content 10.6%, ash content 5.6%, nitrogen content 9.6% content): Same M-5 (moisture content 9.5%) %, ash 4.5%, nitrogen 7.8%); same M-6 (moisture 9.2%, ash 4.7%, nitrogen 8%);
．． 4%); Same M-8 (moisture 10.1%, ash 4.6
%, nitrogen content 8.7%) - the same M-9 (moisture 9.8%, ash content 4.8%, nitrogen content 8.5% content) The mice (0.1 to 5 g/kg) and rats (2 to 8 g/kg) were individually dosed by gavage with a throat prodder in increasing doses. During the test period, animals were kept at an animal room temperature of 22-23°C.
The mice were maintained at No deaths were observed at all drug doses administered. Toxicity and behavior after administration were observed over time, but no differences were observed from the normal animal group. Furthermore, there was almost no difference in weight gain from the normal animal group. A post-test necropsy revealed no comparable macroscopic damage to any major organs. Therefore, earthworm dry powder LD, due to its very low toxicity. The value could not be determined.

A-1. Acute toxicity test of earthworm extract: Measured by the same method as the acute toxicity test method in A above. That is, weight 3
An acute toxicity test was conducted by oral administration using 0±2 g of ddy male base mice in each group of 5 animals.

For example, the above-mentioned earthworm extracts of the present invention X-■, -■,
-■, -■, -■, -■, -■, and -■ were administered to the above-mentioned mice at increasing doses from 0.1 g/kg to 5 g/kg using a throat probe. Individually dosed.

Mice were maintained at an animal room temperature of 22-23° C. during the study and observed for 14 days after dosing. No deaths were observed at the doses administered. Moreover, no abnormalities were observed in the findings of poisoning symptoms or in the autopsy. Therefore, the extracts from earthworms of the present invention with aqueous and/or organic solvents are LD, due to very low toxicity. The value could not be determined.

A-2. Acute toxicity test of mixture of earthworm extract and earthworm dry powder: Measured by the same method as in A above. That is, the weight is 30
An acute toxicity test was conducted by oral administration using 5 animals in each group of ddy male bases weighing ±2 g.

A 50:50 (wt%) mixture of the earthworm extract X-■ obtained in the above production methods 3 and 4 and the earthworm dry powder and -1; -2; X-
■Toni-3; X-■Toni-7, X-■-1 Toni-7
;X-■,! :ni-9;X-@lni-7;X
- ■ Using 9 kinds of 50:50 (it%) mixtures of Toni-8, the above mice were injected with O, Ig/kg to 5 g/kg.
kg, and administered individually by gavage with a throat prodder. Mice were maintained at an animal room temperature of 22-23° C. during the study and observed for 14 days after dosing. No deaths were observed at the doses administered. Moreover, no abnormalities were observed in the findings of toxic symptoms or autopsy. Therefore, the mixture of aqueous and/or organic solvent extracts from earthworms and earthworm dry powder of the present invention has LD, due to its very low toxicity. The value could not be determined.

B. Pharmacological test of the effect of dried earthworm powder on experimental diabetic mice: 1) Animals: 1 group of ddy male mice weighing 30±2 g 5
The fish were used.

2) Feed and rearing conditions: Mice were placed in one cage using solid feed manufactured by Nippon Talea, and feed and water were available ad libitum. The mice were raised at constant temperature and humidity at a temperature of 23±1° C. and a humidity of 55±5%.

After fasting the DDY male mice for 16 hours, 75 mg/kg of alloxan was administered intravenously, and 48 hours later, earthworm dry powder M-1 (moisture 1O92%, ash 5.1%,
Nitrogen 9.4%), same M-2 (moisture 1O94%, natural content 5.
3%, nitrogen 8.6%), M-3 (moisture 1007%, ash 5.2%, nitrogen 9.2%), M-4 (moisture 10.6%),
%, ash 5.6%, nitrogen 9.6%), same M-5 (moisture 9
．． 5%, ash 4.5%, nitrogen 7.8%), M-6 (moisture 9.2%, ash 4.7%, nitrogen 8.4%).
00 mg/kg) was suspended in physiological saline and administered orally, blood was collected from the heart 150 minutes later, and the amount of sugar in the blood was measured by the glucose oxidase method.

The measurement results are shown in Table-5. Dry earthworm powder was found to significantly lower blood sugar.

* : p < 0.05, * * : P < 0.01,
** *: P <0.001 B-1, Pharmacological test 2 of the effect of earthworm extract on experimental diabetic mice: Measured by the same method as the pharmacological test of the effect on experimental diabetic mice in B above. i.e. l) Animal double weight 3
One group of 5 ady male mice weighing 0±2 g was used.

2) Feed and rearing conditions: Solid feed manufactured by Nippon Yurea Co., Ltd. was used, one mouse was placed in each cage, and feed and water were allowed ad libitum. The mice were raised under constant temperature and humidity conditions at a temperature of 23±1° C. and a humidity of 55±5%. After fasting for 16 hours, 75 mg/kg of alloxan was administered intravenously to the above ddy male base, and 48
After an hour, four types of earthworm dry powder (K-1, -3, -7
and -9] at 300 mg/kgs, and nine types of earthworm extracts of the present invention [X-■, -■, -■, -■, -■, -■,
-[Phase], -■ and -■] were each suspended in physiological saline and administered orally at 50 mg/kg. After 150 minutes, blood was collected from the heart and the blood sugar level was measured by the glucose oxidase method.

The measurement results are shown in Table-5-1. It was found that the dried earthworm powder and earthworm extract of the present invention significantly lower blood sugar. The earthworm extract had a greater reduction in blood sugar levels than the dried earthworm powder at an amount of 176 times the dried earthworm powder.

(Margin below) Table-5-1 *: P<0.05, * *: P<0.01. ＊ ＊
*+P<0.001 B-2, Pharmacological test 3 of the effect of the mixture of earthworm extract and earthworm dry powder on experimental diabetic mice: Measured by the same method as the pharmacological test of the effect on experimental diabetic mice in B above. i.e. 1) Animal double weight 30
One group of 5 ddy male mice weighing ±2 g was used.

2) Feed and feed conditions Nippon Talea Co., Ltd. hardened feed was used, one mouse was placed in each cage, and feed and water were allowed ad libitum. The animals were kept at a constant temperature of 23±1°C and humidity of 55±5%. After fasting for 16 hours, 75 mg/kg of alloxan was administered intravenously to the ddy male base, and 48 hours later, 15 mg/kg of each of the 10 kinds of earthworm extracts and 90 mg/kg of each of the 10 kinds of dried earthworm powder were administered. (The combinations of this mixture are shown in Table 5-2.) and three types of earthworm extracts as control drugs [X-■, -〇 and -@
] and 5 types of earthworm dry powder [M-2
, -4, and -6, K-1 and ni-7], 300 mg each
/kg each was suspended in physiological saline and administered orally,
Blood was collected from the heart after 150 minutes, and the amount of blood sugar was measured by the glucose oxidase method. The measurement results are shown in Table-5-3. All of the above subjects were found to significantly lower blood sugar. In particular, a mixture of earthworm extract and earthworm dry powder has a greater reduction in blood sugar levels than earthworm extract and earthworm dry powder alone when compared on a unit weight basis, and the synergistic effect of this mixture is clear.

(Margins below) Table-5-3 *: P < 0.05, * *: P < 0.01, **
*: P<0.001C, Two oral administration experiments of earthworm dry powder to humans Five volunteers who consented to the experiment were administered Capsene preparation C (l) prepared in Reference Example 6 below with dietary therapy. Dry earthworm powder (M-3 above, moisture 1007%, ash 5.2%) in capsules
Capsules A manufactured in Containing Side 4 (containing 150 mg of earthworm dry powder (M-2, moisture 1O64%, ash 5.3%, nitrogen 8.6
%) containing 150 mg] and the capsule prepared in Example 10. Dried earthworm powder (M-2 above) was added to the DCI capsule.
Contains 150mg] capsules 3 times a day after meals.
The drug was administered orally within 0 minutes. Blood was collected from a 62-year-old man every month after administration, and blood was collected after the drug was administered for 9 months.

A 59-year-old man had blood drawn before taking the drug and 2,3, and 4 months after taking the drug. A 76-year-old woman had blood drawn before taking the drug and 4 months after taking the drug.
A 79-year-old woman had blood drawn before taking the drug and 3 and 6 months after taking the drug. A 61-year-old woman had blood drawn before taking medication, and at 1, 3, 4, and 8 months after taking medication. Blood sugar level was measured by the glucose oxidase method. The results are shown in Table-6.

Table 6 (blank space below) [Note] 9 Ma: After 9 months, otherwise the same meaning Standard blood sugar value (m
g/dQ) is 50-100 before breakfast and 150 2 hours after breakfast.
Below, 50-100 before lunch, 150 or less 2 hours after lunch,
50-100 before dinner and 150 or less 2 hours after dinner.

In all five volunteers shown in Table 6, the fasting blood sugar level before breakfast before drug administration was high.
He was a diabetic patient.

In the case of a 62-year-old man who is considered to have moderate diabetes, an improvement in blood sugar levels was observed two months after administration of the dried earthworm powder of the present invention, and three months after administration, among the six test values, the The value for one item after 2 hours was 185 mg/d (2, which was high, but the value had improved to near the standard value. 4 months after administration, it was 6.
All test values for the items reached standard values. Thereafter, the blood sugar level in item 6 remained within the standard range for 5 months.

A 59-year-old man had 6 test values after 3 months of taking medication.
Standard values were reached, except for one item before lunch, which was slightly higher at 114 mg/d4, and four months after administration, all six items were within the standard range.

For a 79-year-old woman, three months after taking medication, the test values for six items were:
Values 2 hours after lunch and before dinner were 171 and 107 mg/d.
The values of the four items other than Q and the slightly high value improved to within the standard value range, and completely reached the standard value 6 months after administration.

The 76-year-old woman had mild diabetes, but her blood sugar levels in six categories reached standard values four months after administration.

A 61-year-old woman has fairly severe diabetes.

Four months after administering the dried earthworm powder of the present invention, an improvement in blood sugar level was observed in this woman, and after eight months, she was able to improve
The values after hours and before lunch were 197 and 210 mg/d, respectively.
(The four items other than the high value 2 reached the standard value.

The oral administration experiment of this dried earthworm powder to humans was successfully completed and without any side effects. For example, even after long-term administration for 6 to 9 months, there was never any risk of hypoglycemia, which is a drop in blood sugar level below the standard lower limit, and the dried earthworm powder of the present invention is considered to be a safe drug. Understood.

The above results can be summarized as follows. That is,
With oral administration of earthworm dry powder, it was somewhat difficult to lower blood sugar levels in all six categories to standard values in patients with fairly severe diabetes; however, after 8 months of administration, It has been found that it has an improvement effect on 4 items, such as lowering blood sugar levels to standard values. However, in patients with mild to moderate diabetes, blood sugar levels in all six categories were able to be reduced to within standard values from 4 months after administration. It has been found that the dried earthworm powder of the present invention is a safe and excellent hypoglycemic agent and an excellent diabetes treatment and prevention agent.

From these results, it is easy to see that earthworm extract, which is an extract from earthworms, is as safe for humans as dried earthworm powder, and is an excellent diabetes treatment and prevention agent. Similarly, mixtures of earthworm extract and earthworm dry powder in arbitrary proportions are also safe for humans.
Moreover, it can be easily seen that it is an excellent diabetes treatment and prevention agent.

That is, although there is a slight difference in the manufacturing method between earthworm dry powder and earthworm extract, since they are manufactured from the same raw material, it is natural that they exhibit excellent effects. Furthermore, since the earthworm extract is a selective extraction of the active ingredients in the earthworm suspension or its dry powder, it is believed that it is effective in smaller amounts than the earthworm dry powder. The reason for the synergistic effect of the mixture of earthworm extract and dried earthworm powder was unknown, but it was an excellent and completely unexpected effect.

Example 1. Capsule E Earthworm extract (X-■) 30mg Lactose 2
8 Microcrystalline cellulose 47 Mannitol 100 Corn starch 40 Polyvinylpyro, Lydon
2-hydroxypropylcellulose
3 total 250 mg After mixing the ingredients other than hydroxypropyl cellulose in the above formulation well using a fluidized bed granulator, a 5% aqueous solution of hydroxypropyl cellulose was sprayed as a binder, and the mixture was dried at 40°C and made into granules. . Each hard capsule was filled with 250 mg of the granules to produce a hard capsule.

Example 2. Capsule F Granule Earthworm extract (X-■) 30mg Lactose 1
03 Corn starch flour 89 Potato starch flour 22 Tarri
3 Magnesium stearate
Total of 3 250 mg According to the above recipe, the well-mixed powder was used in an extractor to produce granules. Hard capsules were manufactured by filling 250 mg of each of the granules into hard capsules.

Example 3. Capsule G Earthworm extract (X-[phase]) 30m
g Phosphoric acid-hydrogen Calcilla! ,,,, 6
0 Sodium hydrogen phosphate 20 Mannitol 38 Magnesium stearate 2 total 150 mg The above formulations were thoroughly mixed, and each 150 mg of this mixed powder was filled into gelatin capsules to produce capsules.

Example 4. Capsule H Earthworm extract (x-o above) 30 mg Sodium lauryl sulfate Sodium 4-phosphate l Manitol
113 Magnesium stearate
2 total 150mg Mix the above prescribed ingredients well. This mixed powder was filled into gelatin capsules in an amount of 150 mg each to produce capsules.

Example 5. Capsule I Earthworm dry powder (M-2) 75 mg Earthworm extract (X-■ above) 15 mg Sodium lauryl sulfate Sodium 4-phosphate l Manitol
153 Magnesium stearate
250mg in total Mix the above prescribed ingredients well. This mixed powder was filled into gelatin capsules in an amount of 250 mg each to produce capsules.

Example 6. Capsule J Earthworm dry powder (M4) 50mg
Earthworm extract (X-■) 20 mg Sodium lauryl sulfate 2 Manitol 176 Magnesium stearate 2 total 250 mg The above formulations are mixed well. This mixed powder was filled into gelatin capsules in an amount of 250 mg each to produce capsules.

Example 7. Dry earthworm powder (M-1) in capsules 75mg earthworm extract (X-■ above) 15mg sodium lauryl sulfate sodium diphosphate-hydrogen l mannitol
55 Magnesium Stearate
2 total 150mg Mix the above prescribed ingredients well. This mixed powder was filled into gelatin capsules in an amount of 150 mg each to produce capsules.

Example 8. Capsule L Earthworm dry powder (-7 above) 50 mg Earthworm extract (X-@l above) 20 mg Sodium hydrogen phosphate 2 Manitol
76 Magnesium stearate 2 total 150mg Mix the above formulation well. This mixed powder was filled into gelatin capsules in an amount of 150 mg each to produce capsules.

Example 9. Capsule M Earthworm dry powder (-9 above) 70 mg earthworm extract (X-■ above) 15 mg milk
M
70 corn starch flour 69 potato starch flour 22 tal
the law of nature
Magnesium 2 stearate
2 total of 250 mg According to the above recipe, granules were manufactured from the well-mixed powder using an extruder. Hard capsules were manufactured by filling 250 mg of each of the granules into hard capsules.

Example 1 O1 Tablet A Earthworm extract (X-■ above) Manitol hydroxypropoxymethyl cellulose tallow Microcrystalline cellulose Hydrogenated castor oil 0 mg Total 80 mg Tablet B Earthworm extract (X-■ above) 30 mg Cornoxide powder 3° milk
Sugar 35t
Ruri
3 Magnesium stearate
2 total 100mg Tablet C Earthworm extract (X - (!D) 15
mg earthworm dry powder (-7 above) 50 Soluble starch 20 Corn starch 25 Microcrystalline cellulose
silicon 45 oxide
3 Magnesium stearate 2 total 1
60 mg According to the above recipe, the powder was uniformly and thoroughly mixed and used in a tablet machine to produce various tablets.

Example 11 1 kg of live earthworms (red earthworms) lightly washed with water
(approximately 20,000 fish) were placed at a temperature of 15°C in an acidic aqueous solution 4Q with a pH of 6.2 in which l=1 mixed acid of malic acid and citric acid was dissolved.
After leaving for 3 hours to fully excrete the fecal soil in the digestive tract, the live earthworms are thoroughly washed with water to remove dirt, feces, and other impurities adhering to the body surface of the live earthworms.

Next, it is wet-pulverized using a mixer. The obtained earthworm suspension was placed in a tray and frozen at -30°C for 40,119 hours, and then the product temperature was kept at -40°C for 0.000. Freeze-dry for 6 hours under a vacuum of lmmHg, then raise the temperature of the shelf on which the tray is placed to 30°C, Q, vacuum-dry for 6 hours under a vacuum of lmmHg, and then
Next, raise the shelf temperature to 50°C, vacuum dry under Q, 2mmHg vacuum for 10 hours, then raise the shelf temperature to 80°C, and dry under a vacuum of 12mmHg.
280 g of dry earthworm powder product (M-1) was obtained by drying under vacuum for 8 hours.

Example 12 Live earthworms (Hkg of red earthworms) lightly washed with water were placed in an acidic aqueous solution 3a of pH 5.5 containing a mixed acid of 1:1:1 of phosphoric acid, tartaric acid and lactic acid at a temperature of 10°C for 2.5 kg.
After leaving it for a while to fully excrete the feces in the digestive tract,
Wash the live earthworms thoroughly with water to remove dirt, feces, and other dirt adhering to the body surface of the live earthworms.

Next, it is wet-pulverized using a mixer. The obtained earthworm suspension was placed in a tray and frozen at -25°C for 20 hours, then the temperature was lowered to -35°C and the temperature was reduced to 0. Freeze-dry for 7 hours under a vacuum of lmmHg, then raise the temperature of the shelf on which the tray is placed to 28°C, Dry earthworm powder by vacuum drying under a vacuum of 1 mmHg for 10 hours, then vacuum drying at 40 °C for 1.13 hours under a vacuum of 0.2 mmHg, then drying at 78 °C under a vacuum of 1 mmHg for 8 hours. Product CM-2) 27
I got 5g.

Example 13゜1 kg of live earthworms (red earthworms) lightly washed with water
is left in an acidic aqueous solution 2a with pH 5.8 in which malic acid is dissolved at a temperature of 13° C. for 3 hours to fully excrete the feces in the digestive tract. Wash the live earthworms thoroughly with water to remove dirt, feces, and other dirt adhering to the body surface of the live earthworms. Next, it is wet-pulverized using an Ultra Homo Mixer (manufactured by Nippon Seiki Co., Ltd.). Place the obtained earthworm suspension in a tray.
After freezing at 0°C for 30 hours, the temperature was reduced to -30°C.
Freeze-dry for 8 hours under a vacuum of mmHg, then raise the shelf temperature on which the tray is placed to 25°C, vacuum dry it for 7 hours under a vacuum of Q, lmmHg, then raise the shelf temperature to 45°C, O, lmm.
Vacuum dry under Hg vacuum for 12 hours, then reduce the shelf temperature to 80°C.
275 g of earthworm dry powder product (M-4) was obtained by drying for 7 hours under a vacuum of nonHg.

Example 14 Dirt, feces, and other impurities adhering to the body surface of 1 kg of live earthworms (red earthworms) are thoroughly washed with water four times to remove them. Next, the live earthworms were placed in 2.5Q of an acidic aqueous solution with a pH of 5.7 in which a mixed acid of malic acid and lactic acid (1:1) was dissolved at a temperature of 15°C.
Leave it at ℃ for 2.5 hours to excrete the feces in the digestive tract.

Next, after lightly washing the live earthworms, they are wet-pulverized using a mixer. Place the obtained earthworm suspension in a tray.
Freeze for 24 hours at °C.

Next, freeze-dry the product for 7 hours under a vacuum of 1 mmHg at a temperature of -35°C, then raise the temperature of the shelf on which the tray is placed to 22°C.
After drying in vacuum for 10 hours under a vacuum of 1 mmHg, the shelf temperature was raised to 42°C and dried for 15 hours under a vacuum of 0.2 mmHg. 265 g of dry earthworm powder product (M-6) was obtained by drying under vacuum of lmmHg for 7 hours.

Example 15 1.5 g of potassium dihydrogen phosphate was dissolved in 2.5 Q of an acidic aqueous solution with pH 6.0 in which citric acid was dissolved, and 1.5 g of live earthworms (Hkg of Futoearth worms) lightly washed with water were added to this solution.
Leave at 5°C for 2 hours to excrete the feces in the digestive tract. Next, the live earthworms are washed twice with water to remove dirt, feces, straw, and other impurities that adhere to the body surface of the live earthworms. Next, it is wet-pulverized using a mixer.

After that, put the earthworm suspension into a tray and get it.
Freeze at 0°0 for 24 hours. Next, reduce the temperature of the product to -40°C.
Freeze-dry under vacuum at lmmHg for 5 hours, then freeze-dry at 25°C for 0. After drying for 8 hours under a vacuum of lmmHg, then
0 at 5°C. 280 g of earthworm dry powder product (M-7) was obtained by drying under a vacuum of 1 mmHg for 12 hours and then at 80° C. for 7 hours under a vacuum of 1 mmHg.

Example 16: Dirt, feces, straw, and other impurities adhering to the body surface of 1 kg of live earthworms (Tsurimimizu) were washed thoroughly with water 5 times to remove them. Next, the live earthworms were washed with pH 5 solution containing succinic acid.
, 7 acidic aqueous solution 312 (in which sodium acetate Ig
and 0.5 g of sodium sulfate. ) for 2.5 hours at 13°C to excrete the feces in the digestive tract. Next, the live earthworms are lightly washed with water and then wet-pulverized using a homogenizer. Place the obtained earthworm suspension in a tray at -35°C.
Freeze for 24 hours. Next, at the product temperature -35℃, O, lmmH
Freeze-dry for 7 hours under a vacuum of 0.3 g, then raise the temperature of the shelf on which the tray is placed to 22°C. After vacuum drying for 10 hours under a vacuum of lmmHg, the shelf temperature was raised to 42°C and the thickness was 0.2mm.
The product was dried under a vacuum of Hg for 15 hours, and finally the shelf temperature was raised to 78° C. and dried under a vacuum of 1 mmHg for 7 hours to obtain 275 g of a dry earthworm powder product (M-8).

Example 17. Dirt, feces, straw, and other impurities adhering to the body surface of 1 kg of live earthworms (Pterophthalmos) were washed thoroughly with water 5 times to remove them. Next, the live earthworms were placed in an acidic aqueous solution of pH 5.9 (in which 0.7 g of potassium citrate was dissolved) containing 1:1 of a mixed acid of citric acid and tartaric acid.
The feces in the digestive tract were excreted after being left at 5°C for 2 hours. Next, after lightly washing the live earthworms with water, they are wet-pulverized using a blender. Put the obtained earthworm suspension into a tray-356
It was frozen at 0 for 24 hours.

Next, the product temperature is -35℃ and 0. Freeze-dry for 7 hours under a vacuum of lmmHg, then raise the temperature of the shelf on which the tray is placed to 22°C, Q, vacuum-dry for 10 hours under a vacuum of lmmHg, and then
Next, the shelf temperature was raised to 42°C and dried under a vacuum of O-2 mmHg for 15 hours. 283 g of dry earthworm powder product (M-9) was obtained by drying under vacuum of lmmHg for 7 hours.

Example 18゜ Live earthworms (red earthworms) lightly washed with water l kg
Leave it in 3Q of fresh water at a temperature of 10°C for 16 hours to excrete the feces in the digestive tract. Wash the live earthworms thoroughly with water to remove dirt, feces, and other dirt adhering to the body surface of the live earthworms. Next, wet grinding is performed using an ultra homo mixer.

The obtained earthworm suspension is placed in a tray and frozen at -25°C for 15 hours. Next, the temperature was lowered to -35°C and freeze-dried for 6 hours under a vacuum of 11mmHg, and then dried at 30°C for 10 hours under a vacuum of 8mmHg! : Later, 0.2 at 40℃
Dry under vacuum at mmHg for 15 hours, then Q at 78°C.
, 245 g of earthworm dry powder product (M-3) was obtained by drying under a vacuum of 1 mmHg for 8 hours.

Example 19 Live earthworm (red earthworm) 1 kg of dirt, such as mud and straw, adhering to the body surface was washed thoroughly with water 5 times to remove it. Next, the live earthworms were placed in fresh water of 2.5A at a temperature of 15℃.
The feces in the digestive tract were excreted after being left at ℃ for 18 hours. Next, the live earthworms were lightly washed and then wet-pulverized using an ultra homo mixer. Place the obtained earthworm suspension in a tray for 40 minutes.
Freeze at ℃ for 24 hours. Next, the temperature of the product is -40℃, Q, lm
Freeze-dry for 5 hours under mHg vacuum, then at 25 °C.
After drying for 8 hours under +nmHg vacuum, then 45°
O'tlsO, dried under vacuum at lmmHg for 12 hours,
Then at 80℃ 0. 240 g of earthworm dry powder product (M-5) was obtained by drying under vacuum at lmmHg for 7 hours;

Reference Example 16 Tablet A Earthworm dry powder (M-8) 150 mg mannitol 123 hydroxypropoxymethylcellulose microcrystalline cellulose tal Hydrogenated castor oil Tablet B Earthworm dry powder (M-8) Corn starch milk powder Sugar talc Magnesium stearate Tablet C Earthworm Dry Powder (M-8) Soluble Starch Corn Starch Microcrystalline Cellulose Silicon Oxide Magnesium Stearate Total 350 mg 150+og Total 300 mg 50 mg Total 350 mg Mix well and uniformly according to the above recipe; Tablets of various weights were manufactured.

Reference example 2. Granule A Dry earthworm powder (M-2) 150mg lactose
20 Microcrystalline Cellulose 6° Corn Starch 15 Hydroxypropyl Cellulose 5 Total 250 mg According to the above recipe, using a fluidized bed granulator, dry earthworm powder, lactose, microcrystalline cellulose, and corn starch were thoroughly mixed to form hydroxypropyl cellulose. A 5% aqueous solution of was sprayed as a binder and dried at low temperature to form granules.

Reference Example 3.1 Granule B Dry earthworm powder (M-2) Manitol microcrystalline cellulose carboxymethylcellulose calcium stearin 00mg 1,5 Hydrogenated oil 1,5 Total 20 0,0mg Granule C Dry earthworm powder ( M-2) 150mg lactose
53 Corn starch flour 39 Potato starch flour 2 tal
the law of nature
3 Magnesium stearate
A total of 250 mg of the three well-mixed powders were used in an extruder to produce granules according to the above recipe.

Reference example 4. Capsule A Dry earthworm powder (M-2) Lactose Microcrystalline Cellulose Manitol Corn Starch Polyvinylpyrrolidone Hydroxypropylcellulose 50mg Total 250mg In the above formulation, ingredients other than hydroxypropylcellulose may be added using a fluidized bed granulator. After mixing, a 5% aqueous solution of hydroxyglobil cellulose was sprayed as a binder, and the mixture was dried at low temperature to form granules. Hard capsules were prepared by filling 250 mg of each of these granules into hard capsules.

Reference example 5. Capsule B Granules C manufactured according to Reference Example 5 were put into hard capsules, 25
Hard capsules were prepared by filling 0 mg each.

Reference example 6. Capsule C Dry earthworm powder (M-4) 150mg Calcium hydrogen phosphate 60 Sodium hydrogen phosphate 10 Manitol
28 Magnesium Stearate
250mg in total Mix the above formulations well and add this mixed powder to No.
Capsules were prepared by filling 250 mg of each gelatin capsule into 1 liter gelatin capsules.

Reference example 7. Enteric-coated tablet earthworm dry powder (M1) 100mg
Manitol 1O Microcrystalline Cellulose 85 Calcium Carboxymethyl Cellulose 2 Magnesium Stearate 1.5 Total 200 mg After producing plain tablets from the uniformly mixed powder using a tablet machine according to the above recipe, use the following enteric-coated skin coating agent. Enteric coated tablets were prepared.

Coating agent hydroxypropyl methyl cellulose phthalate 14.
8mg Dioctyl phthalate 2.3 Stearic acid 2.3 Light silicon oxide 0.6 Total 20.0mg Reference example 8. Powder A Earthworm dry powder (M-4) 150mg Manitol 50 Corn starch 50 Total 250 mg Powder B Earthworm dry powder (M-4) 150 mg Calcium phosphate-hydrogen 20 Corn starch 80 Total 250 mg The above ingredients were placed in a conical mixer. The mixture was mixed well and uniformly to form a powder.

Reference example 9. Suppositories A Dry earthworm powder (M-5) 200mg Witegsol 540 (Witep
sol) E-85 Uitepsol 1.454 (//
) W-35 Methyl parahydroxybenzoate 3 Butyl para hydroxybenzoate 3 total 2,200 mg Fully open B Earthworm dry powder CM-6) 200 mg Butyl hydroxyanisole 6 total 3.106
mg A melt obtained by thoroughly mixing each of the above-mentioned formulations was poured into an aluminum mold, and the mixture was cooled to produce a full opening.

Reference example 10. Capsule D Earthworm dry powder (M-2) 150mg Sodium lauryl sulfate Sodium 4-phosphate l Manitol
93 Magnesium Stearate
250mg in total Mix the above prescribed ingredients well. This mixed powder is No.
, 1 gelatin capsules were filled with 250 mg each to produce capsules.

〔Effect of the invention〕

As stated above, the present invention relates to a therapeutic agent for diabetes containing an earthworm extract or a mixture of an earthworm extract and earthworm dry powder as an active ingredient. By administering earthworm extract, a mixture of earthworm extract and dry earthworm powder, or dry earthworm powder to experimentally diabetic mice induced by alloxan, blood sugar levels could be significantly lowered.

Next, five diabetic volunteers were given one dose of earthworm dry powder capsules (150 mg content) along with their diet.
The tablet was administered orally three times a day after meals for 4 to 9 months. At least 1 month after 1 month, 2 months, or 3 to 4 months after administration, blood was collected 6 times before breakfast, 2 hours after breakfast, before lunch, 2 hours after lunch, before dinner, and 2 hours after dinner. The amount of medium sugar was measured. As a result, in patients with mild to moderate diabetes, improvement occurred 2 to 3 months after administration of dry earthworm powder, and after 4 months of administration, blood sugar levels for all six items listed above were reduced to standard values (
The standard value of blood sugar before three meals (breakfast, lunch, and dinner) is 50 to 100 mg.
/d12. The standard blood sugar value 2 hours after each of the same three meals is 150.
mg/dQ or less). In the case of severe diabetic patients, it was difficult at best to lower the blood sugar levels of all six items above to within the standard range, but after 8 months of administration, the improvement was achieved by lowering the blood sugar levels of 4 of the 6 TX items to standard values. It was shown to be effective. Also, earthworm dry powder 6
Even during long-term administration for ~9 months, there was no risk of hypoglycemia, where blood sugar fell below the lower limit of the standard value.

Animal experiments using earthworm extracts, mixtures of earthworm extracts and dry earthworm powder, and dry earthworm powders, as well as oral administration experiments of earthworm dry powders to humans, revealed that the earthworm extracts, earthworm extracts, and dry earthworm powders of the present invention The mixture is safe;
It was also found to be an excellent diabetes treatment and prevention agent.

Patent applicant: Co., Ltd., I, I1.・De") Procedural formalities (spontaneous) i, Display of 4 cases 1999 Patent Application No. 12037 2, Title of invention: Antidiabetic agent 3, Relationship to case Patent applicant postal code: 880 Address: Miyazaki 5-1-23 Tachibana Dori Nishi, Miyazaki City, Prefecture
5. There is no increase in the number of claims due to the amendment. 6. "Claims J" and "Detailed Description of the Invention J" in the specification subject to the amendment
W.J.

Attachment (1) The scope of patent claims will be amended as follows.

2. Claim 1: The active ingredient is an earthworm extract obtained by extracting earthworms 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. Diabetes treatment agent containing as.

2. Earthworms smell raw earthworms with organic acids, inorganic acids, organic acid sodium salts, inorganic acid sodium salts, organic acid potassium salts, and! l! After leaving the worms in an aqueous solution containing 0.3% (by weight) or less of at least one compound selected from the group consisting of organic acid potassium salts or in fresh water to remove the excrement in the digestive tract of the live worms, An earthworm suspension obtained by washing and removing dirt adhering to the body surface with water and performing wet crushing, or a live earthworm obtained by washing and removing dirt adhering to the body surface of live earthworms with water, using the above-mentioned aqueous solution history or A claim consisting of an earthworm suspension obtained by leaving the living earthworms in fresh water to remove the excrement from their digestive tracts, then washing and removing the filth adhering to the body surface of the live earthworms with water, and performing wet crushing. 1. The antidiabetic agent according to 1.

3. Earthworms consume the earthworm suspension according to claim 2 from -10 to -
After freezing at 60℃, the temperature is raised stepwise within the range of -60 to 80℃ and the vacuum level is 10 to
Freezing and vacuum drying were carried out for 100 hours, during which the final step of vacuum drying was carried out at 70 to 80°C with a concentration of 0.01 to [l, 5
The antidiabetic agent according to claim 1, comprising dried earthworm powder obtained by drying under a vacuum of mmHH for 5 to 10 hours.

4. A therapeutic agent for diabetes containing a mixture of earthworm extract and dry earthworm powder as an active ingredient.

(2) “External” in line 11 on page 3 of the specification is “extracorporeal.”
I will correct it.

(3) "No. 24" in line 2θ on page 4 of the specification is r24
I corrected it to ``Volume''.

(4) Line 13 on page 14 of the specification, line 17 on page 18
In line 10, page 19, line 10, and page 41, line 4, ``first period'' is corrected to ``1 above''.

(5) "Manufacturing" in line 13 on page 14 of the specification will be corrected to "r manufacturing method."

(6) The amino acid "tyroline" in line 17 of Table 3 on page 23 of the specification is corrected to "tyrosine 1."

(7) Line 15 on page 31 of the specification and line 5 on page 32
row r Coelonic” to rcelonic
First, I corrected it to J.

(8) “Succinic acid or” on page 33 of the specification, line 17
Correct it to ``r-succinic acid, malic acid, or''.

(9) "S-Butyl" in line 8 on page 34 of the specification will be corrected to rl-butyl, S-butylj.

(10) “Vinegar!” on page 35 of the specification, lines 10-11
First, correct "2-" to "acetic acid 2-".

(11) "About 30℃" in the second line of page 44 of the specification is r
Correct it to approximately 1 at 30℃.

(12) On page 44 of the specification, line 4: “-day and night,
” should be corrected to “-Leave it undisturbed day and night.”

(13) “Optionally” in line 16 of page 64 of the specification is replaced with “
I am corrected to say "any."

(14) In the second line of page 66 of the specification, "Hard capsule granules" is corrected to "Hard capsule J for granules."

(15) "Corn" in line 20 on page 70 of the specification is corrected to "corn."

(16) “Reference Example 5” on page 82, line 9 of the specification is changed to “
Reference example 3" is corrected.

(17) rM-4, on line 12 of page 82 of the specification, is r
I will correct it to M-3J.

(18) rM-5, on line 13 on page 84 of the specification, is r
I will correct it to M-9J.

Patent applicant: Eimei Co., Ltd.

Claims (1)

[Claims] 1. An active ingredient is an earthworm extract obtained by extracting earthworms 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. Diabetic treatment agent containing as. 2. When earthworms feed raw earthworms with at least one compound selected from the group consisting of organic acids, inorganic acids, sodium salts of organic acids, sodium salts of inorganic acids, potassium salts of organic acids, and potassium salts of inorganic acids, 0.3 (by weight) Earthworm suspension obtained by leaving the living earthworms in an aqueous solution or fresh water containing less than 20% of the total feces to remove the excrement from the digestive tract of the live earthworms, and then washing away the dirt adhering to the body surface of the live earthworms with water and performing wet crushing. After removing the solution or dirt adhering to the body surface of the live earthworm with water, the live earthworm is left in the aqueous solution or fresh water to remove the fecal soil in the digestive tract of the live earthworm, and then the body surface of the live earthworm is washed with water. 2. The therapeutic agent for diabetes according to claim 1, which comprises an earthworm suspension obtained by washing and removing adhering dirt with water and performing wet grinding. 3. Earthworms consume the earthworm suspension according to claim 2 from -10 to -
After freezing at 60°C, the temperature was raised stepwise within the range of -60 to 80°C and the vacuum level was 10 mmHg or less.
Freeze and vacuum dry for ~100 hours, including the final step of vacuum drying at 70 to 80°C to a thickness of 0.01 to 0.5 mm.
The antidiabetic agent according to claim 1, which comprises dried earthworm powder obtained by drying under Hg vacuum for 5 to 10 hours. 4. A therapeutic agent for diabetes containing a mixture of earthworm extract and dry earthworm powder as an active ingredient.