JPH0651633B2 - Antidiabetic - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention relates to a therapeutic agent for diabetes containing OK-432 (trade name Picibanil, a product of Chugai Pharmaceutical Co., Ltd.) as an active ingredient.

OK-432, which is a streptococcal preparation, is widely used as a therapeutic drug for various cancers such as digestive organ cancer and lung cancer. In addition, it has recently been revealed that this drug exerts tumor resistance through its effects on host functions such as γ-interferon producing ability and NK cell activity. (Celluler Immunol
(Ogy, 1982, 68: 187-192, JNCI, 1982, 69: 807-812) However, nothing is known about the efficacy of this drug for diabetes.

As a result of intensive studies on a drug for treating diabetes, the present inventor has found that the drug is effective for NOD (spontaneous lean diabetes) mice. The present invention has been completed through further research based on this new finding.

OK-432 used in the present invention is a bacterial cell preparation obtained by treating Streptococcus pyogenes (Group A type 3) Su strain (ATCC21060) in the presence of penicillin G under certain conditions and freeze-drying. Sold as a tumor agent.

The drug of the present invention can be administered orally or as an injection. The dose is usually about 0.5 mg to 60 mg as a freeze-dried powder for an adult per day, and about 0.02 mg to 5 mg in terms of dry cells. The preferred dose is 0.02 mg to 1 mg per dry cell when used as an injection, and 0.5 mg to 10 mg per dry cell when used as an oral preparation. The dose of the drug of the present invention can be appropriately increased or decreased depending on the symptoms. The drug of the present invention is usually used by dissolving it in physiological saline or glucose.

[Drug action]

The effect of the preparation of the present invention on diabetes can be confirmed by preventing the onset of spontaneous onset lean diabetic mouse (NOD) mouse.
The experimental method and results are shown below.

(Method) 34 female NOD mice were divided into two groups according to age.
In experiment (1), each group consisted of 7 mice,
Once a week from 4 weeks to 24 weeks, 2K.E of OK-432 (0.2 mg as dry cells) and physiological saline were intraperitoneally administered to the control group once a week. In experiment (2), 10 mice were also OK-432.
Saline was administered to 10 mice in the control group.

Mice in which urine sugar was continuously confirmed for at least one week were sacrificed as diabetic mice, and all surviving mice were sacrificed at the end of the experiment. The sacrificed mouse pancreas was examined histologically.

The pancreas sections of the killed mice were examined histologically. The pancreatic islet inflammation of each pancreatic islet was divided into 4 grades based on the histological evaluation. Grade 0 has no lymphocytes on La Isla Island, Grade 1 has 25% or less of the area of infiltration of lymphocytes, 25 to 50% for Grade 2 and 50 for Grade 3.
% And above.

The blood glucose level was measured by the glucose oxidase method.

(Results) In the experiment (1), 7 female mice to which OK-432 was administered did not develop diabetes up to 24 weeks of age, whereas 6 out of 7 mice to which saline was administered were the same. Diabetes developed during the period (see Figure 1).

In the experiment (2), the OK-432-administered group did not develop diabetes until the age of 24 weeks, whereas 8 out of 10 cases in the control group developed diabetes (see FIG. 2).

The blood glucose level at satiation was 102 ± at the age of 24 weeks in the OK-432 administration group.
It was 18 mg / dl (n = 5, random sampling), whereas in the mice treated with physiological saline, at the time of onset of diabetes.
511 ± 82 mg / dl (n = 5, random sampling).

As a result of histological examination, OK-432 protected the pancreatic islets from lymphocyte infiltration (see Table 1). As is clear from Table 1, in the NOD mice treated with OK-432, 98% of the islets of Lae were normal (grade 0; 38%) or slightly infiltrated with lymphocytes (grade 1; Grade 1; 60%), whereas in the control group, all the islets at the time of onset of diabetes were associated with severe islet inflammation (grade 2 or 3).

[Toxicity] Acute toxicity of the drug of the present invention [LD50 (dry cell weight / kg)]
Are shown in Table 2 below.

Reference example Streptococcus pyogenes (Group A type 3) Su strain (ATC
C21060) was statically cultivated at 37 ° C for 24 hours and the resulting culture solution
l 5% yeast extract medium 1 (yeast extract 50 g distilled water 70
Dissolve in 0 ml, adjust to pH 7.4 with sodium hydroxide,
After heating and cooling at 0 ° C. for 60 minutes, the mixture is passed and the solution is made up to 1 with distilled water. Put this in a sterile flask at 1 kg / cm ²
10 minutes steam sterilization) and incubate at 37 ° C for 20 hours. The culture solution is subjected to centrifugation to collect the cells, 50 ml of cold physiological saline containing 1% hydrogen peroxide is added to the cells to form a suspension, and the suspension is kept under ice cooling for 30 minutes. Next, this suspension is spun down in the cold state, physiological saline is added to the cells, the cells are resuspended and then spun down, and the same operation is repeated once for the cells.

The obtained bacterial cells were suspended in cold BBM (Bahnheimer's basal medium), the absorbance at 660 mμ was measured, and the dry bacterial cell amount was calculated from the value to adjust the BBM liquid amount to 6 mg / ml ( Total liquid volume 60 ml). A solution prepared by dissolving penicillin G potassium in physiological saline in 50 ml of this suspension (1.6 × 10 ⁵ units / ml)
10 ml of the mixture is added, and the mixture is kept at 37 ° C. for 20 minutes and further at 45 ° C. for 30 minutes and then immediately ice-cooled (as dry cell amount 5 mg / ml).
To this solution, 60 ml of an ice-cooled 1% DL-methionine aqueous solution containing penicillin G potassium (penicillin G potassium, 1.08 × 10 ⁵ units / ml) was added, and each 2 ml was dispensed into a 5 ml vial bottle and freeze-dried to prepare a powder formulation. And This dried preparation contains 1 mg / ml of dry cells when reconstituted with 5 ml of physiological saline.

[Brief description of drawings]

1 and 2 show the effect of OK-432 on the development of diabetes in NOD mice.

Claims (1)

[Claims] 1. A therapeutic agent for diabetes containing OK-432 as an active ingredient.