JPS5959623A - Antidiabetic agent - Google Patents

Abstract

PURPOSE:To provide a drug containine humulong or lupulone obtained by extracting hop with a specific solvent as active component, capable of lowering the blood sugar level, and useful as an antidiabetic agent having low side effects. CONSTITUTION:The agent contains humulone or lupulone as active components. The components can be prepared as follows. The granules of hop are extracted with an alcohol, the extract is evaporated to dryness, and the residue is extracted with ether. The extract is treated in similar manner successively with alcohol and n-hexane, and the residue is dissolved in alcohol. Alcoholic lead acetate is added to the solution, the precipitate is filtered and dissolved in alcohol, the solution is made acidic with hydrochloric acid, filtered, and the filtrate is extracted with petroleum ether to obtain humulone. The filtrate obtained after the addition of the alcoholic lead acetate is concentrated, washed with saline water, and extracted with petroleum ether to obtain lupulone. The agent can be administered orally without causing hypoglycemic phenomenon.

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention relates to an antidiabetic agent containing Tumurone or Ruburon as an active ingredient.

Tumron and Lebron contain hop cones, for example, in the 'Journal of American Chemistry'.
Society WJ Vol. 73, pp. 54652-4654 (
1951) and is well known.

The present inventors have discovered that Tumuron or Ruburon are effective as antidiabetic agents, and have completed the present invention.

In addition, the present inventors have also discovered a new method for extracting Tumron or Rubron from hops, which are effective as antidiabetic agents, by the method described below.

That is, the present inventors have found that Tumurone or Leburon obtained from hops by the extraction method using a specific solvent according to the present invention is effective as an antidiabetic agent. 0 Hops have traditionally been mainly used for beer brewing. However, there are still no reports of specific cases in which hop organic solvent extract components, particularly Tumurone or Leburon, are effective against diabetes. The present inventor developed experimental diabetes in experimental animals and examined its suppressive effects, and while variously examining the effectiveness of crude drugs and Chinese herbal medicines,
The present invention was achieved by discovering that Tumuron or Ruburon obtained from hops in a specific solvent exhibits a characteristic and strong anti-diabetic effect.

According to the present invention, it has been found that the method θ is effective in obtaining tumuron or ruburon from hop granules.

First, hop granules are extracted with alcohol, the alcohol is removed by evaporation from the obtained extract, the obtained residue is extracted with ether, the ether is removed by evaporation from this extract,
The obtained residue was further extracted with alcohol, the alcohol was removed by evaporation from the extract, and the residue was then treated with n-hexane→non-n. From the n-hexane solution obtained by r separation, n-
The hexane was removed by evaporation, the residue was dissolved in alcohol, a lead acetate alcohol solution was added to the alcohol solution to form a precipitate, the precipitate was filtered out, the precipitate was dissolved in alcohol, acidified with hydrochloric acid and filtered. , R liquid is extracted with petroleum ether, the petroleum ether layer is separated, and the petroleum ether is removed by evaporation to obtain an antidiabetic agent containing Tumuron as an active ingredient.

In addition, the above lead acetate alcohol solution was added to form a precipitate, the precipitate was separated, the resulting ρ solution was concentrated, washed with brine, extracted with stone 2111 ether, the petroleum ether layer was washed with water, and then the petroleum ether layer was washed with water. By distilling off, an antidiabetic agent containing Ruburon as an active ingredient is obtained.

The method for fractionating active ingredients according to the present invention is a conventionally commonly used extraction process, but is characterized by the fact that active ingredients can be extracted more efficiently using a specific extraction solvent.

The extraction process in the present invention aims to remove impurities unrelated to anti-diabetic effects from hops and extract an anti-diabetic agent containing Tumuron or Rubron, which has stronger medicinal effects, as an active ingredient. It is.

If the purpose is to obtain an extract containing anti-diabetic substances from hops, it is possible to achieve this purpose using conventionally commonly used alcohols and water, but such extracts do not have anti-diabetic effects. The content of components unrelated to this increases, and if the basolateral substance is used as it is, it cannot be expected to have a strong enough effect to be used as an antidiabetic agent. Therefore, there is significance in using a specific extraction solvent as in the present invention.

However, even if the extract contains a high proportion of the above-mentioned medicinal substances unrelated to medicinal effects, if it is treated as a causative agent and extracted with a specific extraction solvent, it can be considered as the extracted extract of the present invention. can.

Hereinafter, the present invention will be explained in more detail with reference to production examples, pharmacological tests, acute toxicity tests, formulation examples, etc.

Production example 1 Add 50% methanol, which is 5 times the amount of herbal medicine, to dried hops,
In the case of cold maceration, leave it at room temperature for 3 days, repeat the process of filtering 3 times, combine the extracts and concentrate under reduced pressure at below 50℃, and in the case of digestion, leave it at 80-95℃ for 3 times. Repeat the extraction procedure three times and repeat the same procedure as described above to obtain a 50% methanol-dried extract with a yield of 20-25%.
I got it. Next, 5 times the amount of ether was added to a portion of the 50% methanol extract obtained here, digested for 3 hours, and filtered three times.The extracts were combined, and the solvent was distilled off to obtain ether. Extract (approximately 20~ from 50% methanol extract)
To this ether extract obtained 25%), cold methanol was added in an amount 5 times the amount of the extract, and the solvent was distilled off under reduced pressure at a temperature below 30°C, using the methanolite dissolved fraction as the total hops resin fraction. The resin fraction contained approximately 8% of the 50% Metamer extract.

For the total pop resin fraction, add n-hexane 5 times the amount of the fraction, and perform extraction for about 3 hours in a low-temperature oven at 40 to 50°C. was distilled off, the n-hexane soluble part was made into a soft resin fraction, and the n-hexane insoluble part was made into a hard resin fraction.
Approximately 40% and 60% of the soft and node resin fractions were obtained from the total hop resin fraction, respectively.

Dissolve the above soft resin fraction 32 in 25 d of methanol, add 18% lead acetate methanol solution, collect the resulting precipitate, dissolve only the precipitate in methanol and add 1% HCI.
After adding methanol solution and filtering, extract with petroleum ether, wash the petroleum ether layer with 10% Na1L water, and remove the petroleum ether layer! ) (Drying with magnesium hydroxide sulfate yielded crude Humulon 450 kg.

In addition, the supernatant obtained by adding lead acetate to 25η? 5 to e
Concentrate under reduced pressure at below 0°C and add 500 me of 10% NaCl water.
Wash with petroleum ether and extract with petroleum ether layer 100m
After washing all of e with water, drying with anhydrous magnesium sulfate and concentrating under reduced pressure to remove petroleum ether, crude Lubron 600
I got ■.

1. Streptocytosine Pharmacological test on diabetic rats Eight to ten male Wiatar rats weighing 180 to 2001 were fed for 24 hours, and then streptocytosine 43/9 (dissolved in citrate buffer of pH 4,5) was administered for 24 hours. ,
A dose of 2 CC/Ky) was administered into the tail vein, and on the second day after streptocytosine administration, only those who clearly developed hyperglycemia were selected, and the test drug was administered on the next day. Blood was collected at 1.3 and 6 hours, and the blood sugar level was measured to determine the effect. As for the blood sugar purchasing method, blood was collected from the orbital vein (1100 L/=) and the blood sugar level was measured.

As the measurement method, the glucose oxidase-peroxidase method was used in all cases below.

After drug administration, the effectiveness was tested in comparison with the streptocytosine treatment group. In addition, neither feed nor water was given for 6 hours during the evaluation period.

2. Pharmacological test on alloxan diabetic rats Using the same rats as in 1, after 24-hour feeding, alloxan 1 permanent 30'mg/Kg (PH3(7) HCI-dissolved in saline solution, dosage volume 1it 2 CC/K9) ) was administered intravenously into the tail vein. Only those in which hyperglycemia developed in the second molar after administration of alloxan were selected, and the test drug was administered the next day in the same manner as in experiment 1 below to determine the effect. Rino Wis
A group of 8 male Tar rats were subcutaneously administered with 1001 g/Ky of epinephrine hydrochloride, and at the same time, the test drug was administered. Blood was collected at 0, 1.3, and 5 hours after drug administration, and blood sugar levels were measured.

4. Effect on blood lactic acid Wistar type Mf with a body weight of 180-200 kg
The same procedure as in 1 was conducted using four 17i:Y rats, and the blood lactate level as well as the blood sugar level was measured. The Barker-Summerlan method was used to measure blood lactate levels.

5. Glucose Tolerance Test Method in KK Mice: 1 trx:s of KK male mice, around 282 years old, were fed for 3 hours, then glucose 5f/Ks+ was orally administered. 15 minutes later, each test drug was administered. administered.

At 0, 1.3, and 5 hours after glucose administration, 100 metts of blood was collected from the orbital vein, and the blood sugar level was measured. Note that dd-K male mice (body weight around 282 kg) were used as normal mice.

(3. Glucose tolerance test in dd- mice. Groups of 8 dd-K male mice weighing around 281 kg,
The experiment was carried out in the same manner as in the method for testing glucose tolerance in KK mice in Section 5.

7. Glucose tolerance test material XM in Wistar male rats
One group of 6 male Wistar rats around 200 f.
The study was conducted in the same manner as in the glucose tolerance test method using 5 KK mice.

8. Acute toxicity test A group of 10 healthy male Wistar rats weighing between 160 and 1809 mm was used to determine the LD value and risk rate using the Litchfield-Wilcochran method based on the number of deaths within one week after administration of the test drug. Confidence limits at 5% were calculated.

Discussion As shown in Table 1, both Tumuron and Lubron lowered blood sugar levels and the onset of action tended to be slower than the comparative drug insulin, but the hyperhypoglycemic phenomenon seen with insulin was not observed at all.

Discussion As shown in Table 2, Tumron and Leburon obtained from hops were found to be effective, similar to the effects in Table 1.

From the above results, the extracted extracts of the present invention, altumuron and ruburon, exhibit antidiabetic effects when administered orally, unlike insulin, and do not cause hypoglycemia unlike insulin and tolubutamide, which is seen in piganide antidiabetic drugs. It is considered to be an excellent drug that does not cause hyperlactatemia.

For treatment, it is estimated that 1 to 2 tablets should be taken at a time according to the formulation example described below, and should be taken 2 to 3 times a day. It goes without saying that any extract of the present invention obtained in the above production examples can be made into tablets according to the formulation examples below.

Formulation example 1 Tumuron or Leburon fraction 50mg potato starch 95 ■ Magnesium stearate 5 ■ 1 tablet 15
0η

Claims (1)

[Claims] 1. An antidiabetic agent containing Tumuron or Ruburon as an active ingredient.