JPH0643331B2 - Aldo-reductase inhibitor - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention relates to an aldose reductase inhibitor comprising goshajinkigan.

In recent years, accumulation of intracellular sorbitol through a polyol pathway, which is a glucose metabolism pathway, has been attracting attention as a cause of various complications in diabetes such as cataract, retinopathy, neuropathy, and nephropathy. The polyol pathway is a metabolic pathway in which aldoses such as glucose and galactose are converted into ketose such as fructose through polyols such as sorbitol and galactitol, and it is clear that they are widely present in various organs throughout the body by immunohistochemical techniques. It has become.

The enzyme that catalyzes the first step in this pathway, the conversion between aldose and polyol, is called aldose reductase,
This enzyme is considered to be the rate-limiting enzyme in the polyol pathway. It has been reported that inhibiting this aldose reductase and reducing the production and accumulation of sorbitol is effective in treating complications in diabetic patients. Therefore, the present inventors, as a result of conducting a study on the aldose reductase inhibitory action for various Kampo formulations, ground yellow, beef knees, wild mushrooms, mountain herbs, Kuramaeko, Sawari, 苓苓,
It was found that a Kampo prescription consisting of peony skin, cinnamon bark, and Fuzi, that is, Goshajinkigan, has an aldose reductase inhibitory action. The present invention is based on this finding, and provides an aldose reductase inhibitor consisting of goshajinkigan which is effective for the treatment of complications in diabetic patients. Goshisha-jinki-gan has a description of its constituent crude drugs, dosage, extraction method, etc. in the classical Chinese medicine formula (saiseikata), and is used for various diseases such as lower limb pain, low back pain, dysuria and frequent urination. However, it has never been known that it has an aldose reductase inhibitory action.

According to the classics, Goshigurenki Maru is 5g of ground yellow, 3g of beef knee, 3g of Yamatake mushroom, 3g of mountain herb, 3g of Mazenko, 3g of Sawara, 3g of Furei.
g, peony skin 3g, cinnamon bark 1g, and adzuki bean paste 1g can be infused with 300ml of water to make 180ml, and this can be taken as an aldose reductase inhibitor, but it is a Chinese herbal medicine that is easy to take and convenient to carry. The extract can also be used as an aldose reductase inhibitor.
For example, 5 to 8 parts by weight of ground, 2 to 3 parts by weight of beef knees, 2 to 4 parts by weight of yamatake mushroom, 3 to 4 parts by weight of mountain herbs, 2 to 3 parts by weight of sardine, 3 parts by weight of sardine, 3 to 4 parts of 茯苓1 part by weight, 3 parts by weight of peony skin, 1-2 parts by weight of cinnamon bark, 0.5-1 part by weight of sardine with 10 times the amount of water when hot extracted, dried and dried to inhibit aldose reductase. Preparation of powdered goshajinkigan dried powder, which is an agent, and powders, granules, tablets, capsules, etc. according to the usual method of manufacturing the preparation by adding excipients, adjuvants, etc. used in usual preparations It can also be used. If desired, this extract is further dialyzed,
You may refine | purify and use it by a conventional method, such as various chromatography.

A specific example of the production of the aldose reductase inhibitor of the present invention is as follows.

Concrete example 5g of ground yellow, 3g of beef knee, 3g of yamatake mushroom, 3g of mountain herb, 3 carshako
g, Sawarei 3g, Furei 3g, Peony skin 3g, cinnamon skin 1g, and adzuki bean 1g mixed with a 10-fold amount, that is, 280ml of water was added and the mixture was heated and extracted at about 100 ° C for 1 hour. It was spray-dried to obtain 3.2 g of dry extract powder.

Next, it will be described with reference to experimental examples that the aldose reductase inhibitor of the present invention has an aldose reductase inhibitory action.

Experimental Example 1 <Measurement of aldose reductase activity> A 6-week-old Wistar male rat was sacrificed under ether anesthesia, and the lens was immediately removed and stored at -20 ° C.

The lens is a 135 mM sodium-potassium-phosphate buffer solution (pH: 0.5 mM phenylmethylfonyl fluoride).
7.0) and homogenize at 30,000 rpm for 3
Centrifuge for 0 minutes. The supernatant was used as a sample for aldose reductase activity measurement. All the above operations were performed at 4 ° C, and the sample was stored at 0 ° C.

The measurement of aldose reductase activity was carried out using Dufram (Dufra).
ne) et al. [Biochemical Medicine, 32, 99-10]
5 (1984)]. That is, 100
mM lithium sulfate, 0.03 mM NADPH (reduced nicot
inamide adenine dinucleotide phosphate) and 0.1 mM DL-glyceraldehyde or 2 as a substrate
The above sample 1 was added to 800 μl of 135 mM sodium-potassium-phosphate buffer (pH 7.0) prepared to contain 0 mM glucose.
00 μl and the drug obtained in the above specific example were added to distilled water at 1 mg / m 2.
100 μl of the drug solution, which was dissolved so as to have a volume of 1, was added to each, and the mixture was reacted at 30 ° C. for 30 minutes. Next, 0.
The reaction was stopped by adding 0.3 ml of 5N hydrochloric acid, and 1 ml of 6N sodium hydroxide containing 10 mM imidazole was added to produce NADP (oxidized form niDP).
Cotinamide adenine dinucleotide phosphate) was converted into a fluorescent substance, and 60 minutes later, the fluorescence intensity was measured.
The fluorescence intensity was measured at room temperature using a spectrophotometer PF-510 (manufactured by Shimadzu Corporation) at an excitation wavelength of 360 nm and a fluorescence wavelength of 46 nm.
It was measured under the condition of 0 nm. Further, the fluorescence intensity measured by reacting in the same manner as above except that distilled water was added instead of the drug solution was used as a control value.

Aldose reductase is a DL-
It is an enzyme that converts glyceraldehyde or glucose into polyol, and NADPH changes into NADP along with this reaction. Therefore, if NADP is low, aldose reductase is inhibited.

As a result, the degree of inhibition of the rat lens aldose reductase activity of the drug obtained in the specific example was 72.6% when the substrate was DL-glycerinaldehyde, and 38.7% when the substrate was glucose. .

From the above results, it was confirmed that the aldose reductase inhibitor of the present invention inhibits the activity of aldose reductase, and is expected to be effective in preventing or treating complications of diabetes.

Next, the acute toxicity test of the oral administration of the drug obtained in the specific example was conducted.
As a result of using Y mice and Wister rats, no drug died even when the drug obtained in the specific example was orally administered at 15 g / kg (administration limit).

Thus, the drug of the present invention has extremely low toxicity and high safety.

Judging from the experimental data and the results of the acute toxicity test in the present invention, the effective dose of the drug of the present invention varies depending on the age, body weight, and degree of disease of the patient. It is considered appropriate to administer the daily dose of 1 to 10 g divided into three times a day according to the symptoms.

Next, the present invention will be specifically described with reference to examples, but the present invention is not limited thereto.

Example 1 200 g of the drug produced according to Example 1 above was added to lactose 95
g and 5 g of magnesium stearate, and the mixture is compression molded, crushed, sized and sieved to 2
Good granules with 0-50 mesh particles were obtained.

This granule has a daily dose of 1.5 to 15g depending on the symptoms.
(Corresponding to 1 to 10 g of the dry extract powder weight of the aldose reductase inhibitor of the present invention) is taken in 3 divided doses.

Example 2 200 g of the drug prepared according to Example 1 above was mixed with 45 g of microcrystalline cellulose and 5 g of magnesium stearate, and this mixture was tabletted with a single-shot tableting machine to produce a tablet having a diameter of 9 mm and a weight of 250 mg. did. One tablet of the present invention contains 200 mg of the drug of the present invention.

The daily dose of this tablet is 5 to 50 tablets 3 to 6 times depending on the symptoms.
Take in divided doses.

Example 3 A hard capsule was filled with 250 g of the drug produced according to Example 1 above. This capsule is 4 times daily depending on the symptoms.
Take ~ 40 capsules 3-6 times daily.

Claims (1)

[Claims] 1. An aldose reductase inhibitor comprising goshajinkigan.