JP2903324B2 - Superoxide scavenger - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION [Industrial Application Field] The present invention relates to a superoxide scavenger which can be used in the fields of medicine, food, cosmetics and the like.

[Conventional technology]

Radical O ₂ occurring at one-electron reduction of superoxide i.e. oxygen molecules ^- are known to act as an important protective factor in vivo. For example, bacteria,
Neutrophils, monocytes,
Phagocytic cells such as macrophages are activated to develop dynamic functions such as migratory ability and phagocytosis, and as a result, lysosomal enzymes and superoxide are produced and secreted, which are directly or thorough in the melting and sterilization of poor foods. Involve indirectly and protect the body from external enemies.

On the other hand, superoxide causes various tissue disorders when present in an excessive amount in a living body. Normally, the amount of superoxide generated in a living body is about 1% or less of oxygen absorbed by respiration, and this is sequentially eliminated by the catalytic action of superoxide dismutase (SOD) contained in cells. In cases where the enzymatic action is reduced, such as in the elderly, the elimination is inadequate and the concentration of superoxide increases, which can lead to tissue disorders such as rheumatoid arthritis and Behcet's disease, and the superoxide or the resulting superoxide. Myocardial infarction, stroke, cataract, spots, freckles, wrinkles, diabetes,
Causes arteriosclerosis, stiff shoulders, and coldness. Even if you are not an old man, skin is an organ that is particularly easy to produce superoxide because it is directly stimulated by environmental factors such as ultraviolet rays,
An increase in the concentration of superoxide and the production of lipid peroxide accompanying the superoxide concentration are apt to occur, which is liable to cause melanin pigment formation, spots, fine wrinkles and the like.

Attempts to use superoxide dismutase as a pharmaceutical or in cosmetics or foods to prevent or treat the various disorders described above due to excess superoxide have been made by heat-labile and inactive enzymes. It has been unsuccessful because of its simplicity and its significant cost.

Attempts to find a substance having a superoxide scavenging action other than superoxide dismutase have been disclosed in
No. 50877, in which baicalein in Huangfen is utilized. However, baicalein is contained only a little in Huangfen, and its extraction is extremely expensive.

[Problems to be solved by the invention]

An object of the present invention is to provide a superoxide scavenger comprising a substance which is not unstable like superoxide dismutase, can be easily incorporated into pharmaceuticals, cosmetics, foods and the like, and is stable and can be obtained at low cost. .

[Means for solving the problem]

The superoxide quencher successfully provided by the present invention is characterized by containing a component extracted from a lower alcohol or a mixture thereof with water from melissa, enmeisou, birch, Nantenmi, kina or Eijitsu. Things.

It is considered that the superoxide scavenger of the present invention scavenges superoxide by a reaction of 2O ₂ ⁻ + 2H ⁺ → H ₂ O ₂ + O ₂ , but it is still unclear what component in the raw material plant scavenges superoxide. Not elucidated.

In order to extract the active ingredient of the superoxide scavenger from the raw material plant, the raw material to be treated is mixed with about 5 to 15 times the amount of a lower alcohol such as methanol, ethanol, isopropanol, or a mixture thereof with water (preferably, water content). Of about 30-80%) at room temperature to elute the soluble components, and after an appropriate time, perform solid-liquid separation. These operations are repeated two or three times, and the obtained extracts are combined and filtered. The obtained extract can be used as it is as a superoxide scavenger, but it is concentrated if necessary, further dried under reduced pressure, and used as a dry extract.

The superoxide scavenger of the present invention can be used in a wide range of applications, such as being used as a pharmaceutical product, and blended into cosmetics (including quasi-drugs), foods, and the like. When blended in cosmetics or the like, the suitable addition amount of the superoxide scavenger of the present invention is about 0.001 to 0.1% in terms of a raw material plant.

〔The invention's effect〕

As described above, the superoxide scavenger of the present invention can be easily produced from a readily available plant by a simple extraction operation and is inexpensive, and its superoxide scavenging action is not impaired by heating,
In addition to being used as a medicament for the prevention or treatment of the aforementioned diseases caused by excess superoxide,
It is very useful because it can be added to cosmetics and the like to promote health and beauty.

〔Example〕

50 v / v% ethanol 100 to 100 g of dried and crushed raw material plant
Add 0 ml and extract at room temperature with occasional stirring for 3 days. The obtained extract is filtered, and the filtrate is allowed to stand at 5 ° C. for 3 days and then filtered again to obtain a clear solution-like superoxide scavenger.

Superoxide scavengers were produced from Melissa, Emmeisou, Birch, Minami Amami, Kina and Agetsu by the above-mentioned production method, and the superoxide scavenging action was examined for them. The test method (NBT method) is as follows.

(1) Reagent 0.05 M Na ₂ CO ₃ buffer (pH 10.2) 3 mM xanthine solution: Dissolve 45.64 mg of xanthine in the buffer to make 100 ml.

3 mM EDTA solution: 11.7 mg of EDTA2Na dissolved in distilled water
Make it 00 ml.

BSA solution: Dissolve 15 mg of Bobvin serum albumin (Sigma) in distilled water to make 10 ml.

0.75 mM NBT solution: Dissolve 61.32 mg of NBT (nitro blue tetrazolium) in distilled water to make 100 ml.

Xanthine oxidase solution: Xanthine oxidase is diluted with distilled water so that the absorbance in the blank test of the operation (2) described later falls in the range of 0.20 to 0.23.

6 mM CuCl ₂ solution: Dissolve 102.29 mg of CuCl ₂ .2H ₂ O in distilled water to make 100 ml.

(2) Operation Transfer 2.4 ml of Na ₂ CO ₃ buffer solution into a test tube, and add 0.1 ml each of a xanthine solution, an EDTA solution, a BSA solution, and an NBT solution. Add 0.1 ml of sample solution (10-fold diluted solution, 50-fold diluted solution, 100-fold diluted solution, 200-fold diluted solution), leave at 25 ° C for 10 minutes,
Add 0.1 ml of xanthine oxidase solution, stir quickly and incubate at 25 ° C. After 20 minutes CuCl ₂ solution 0.1
The reaction is stopped by adding ml and the absorbance is measured at 560 nm. For comparison, superoxide dismutase (Cu
・ Zn-type SOD, activity: 3000-4000unit / mg, Wako Pure Chemicals aqueous solution
The same measurement is performed for 0.1 ml. Separately, a blank test is performed using distilled water instead of the sample.

 Table 1 shows the measurement results.

Further, the superoxide erasing rate of the sample heated at 70 ° C. for 10 minutes was measured in the same manner. Table 2 shows the results.

──────────────────────────────────────────────────続 き Continued on the front page (51) Int.Cl. ⁶ Identification code FI A61K 35/78 ABL A61K 35/78 ABL ABN ABNH ABS ABS ABS ADA ADA ADP ADPQ ADS ADS AED AED (56) References JP-A-60- 224629 (JP, A) JP-A-61-24522 (JP, A) JP-A-60-104005 (JP, A) JP-A-62-205028 (JP, A) JP-A-60-178818 (JP, A) JP-A-2-243632 (JP, A) (58) Fields investigated (Int. Cl. ⁶ , DB name) A61K 35/78 A61K 7/00-7/50

Claims (1)

(57) [Claims] 1. A superoxide scavenger comprising a component extracted from a lower alcohol or a mixture of a lower alcohol and water from melissa, radishes, birch, southern fruit, kina or age.