JPH04334310A - Agent for suppressing formation of lipid peroxide - Google Patents

Abstract

PURPOSE:To provide a safe agent for suppressing formation of lipid peroxide, having low side effect compared with conventional substance and excellent effect for suppressing the formation of lipid peroxide and useful as pharmaceuticals, quasi drugs and cosmetics. CONSTITUTION:The objective effective and safe agent for suppressing the formation of lipid peroxide can be produced by using a lactoferrin as an active component. The amount of the lactoferrin is >=0.001wt.% (preferably >=0.01wt.%) and the agent is coated, rubbed or sprayed (aerosol) to the skin in the form of solution, powder, sol, gel, emulsion, wax and various other forms. It can be used also as an additive for the conventional cosmetic or external agent by conventional process. Since the compound is stable to various other additives and cosmetic bases, it can be used in combination with the substances. Any lactoferrin including those produced by tissue culture or chemical synthesis can be used in the present process provided that the compound has an iron- bonding capability.

Description

Detailed Description of the Invention [0001] [Industrial Application Field] The present invention has the effect of suppressing the production of lipid peroxide, and its application fields include, for example, pharmaceuticals, quasi-drugs, and cosmetics. (people and other movements/
(including various preparations used for plants). [0002] Lipid peroxide is an unsaturated fatty acid, which is a component of lipid, to which peroxide, mainly hydroperoxide, is attached. These lipid peroxides cause damage to cell membranes, inactivation of various enzymes, and denaturation of biological proteins in vivo, and adversely affect biological tissues. These causes are thought to accelerate aging and lead to diseases such as cancerous changes. [0003] In the process of lipid peroxidation, it is necessary to radicalize polyvalent fatty acids or oxygen, and the radicalization is promoted by factors such as light, temperature, and heavy metals. Therefore, substances have been developed that suppress the production of lipid peroxide by removing these factors, preventing radical formation, and decomposing and eliminating generated radicals. For example, radicalized oxygen can be decomposed,
Enzymes and vitamins such as superoxide dismutase, catalase, and glutathione peroxidase that eliminate the enzymes, or plant extracts and coenzyme Q such as quercetin and oryzanol that have antioxidant effects, and SH compounds that decompose by combining with lipid peroxides. Sunscreen agents include pantothenic acid, which suppresses the production of lipid peroxides by promoting albumin and fatty acid β-oxidation, and titanium dioxide and urocanic acid, which suppress lipid peroxidation by blocking ultraviolet rays, which are the main promoting factors. and UV absorbers are known. However, there have been no known examples of applying lactoferrin as a lipid peroxide production inhibitor. Problems to be Solved by the Invention The present inventors have therefore developed a new drug that has fewer side effects (safety) than substances conventionally used as a lipid peroxide production inhibitor, and which suppresses lipid peroxide production. They have been searching for substances with excellent suppressive effects among natural substances. As a result, lactoferrin becomes Fento
The present invention was completed based on the discovery that the compound inhibits the n reaction and suppresses lipid peroxide production. The present invention will be described in detail below. As mentioned above, the peroxidation reaction of lipids begins with the generation of radicals through the reaction of some factor, but transition metals such as cobalt, manganese, iron, and copper quickly undergo hydrolysis. Radicals are generated by decomposing peroxide and hydrogen peroxide. Among these, the decomposition by iron ions is well known as the Fenton reaction, and it also plays an important role in living organisms. [0007] Among iron complexes in vivo, those that can serve as strong initiators of lipid peroxidation are ADP-iron complexes and organic acid-iron complexes, which weakly bind iron. Fen due to Fe2+-ADP complex generated by reduction by enzyme
In a ton-type reaction, hydroperoxide is decomposed into lipid alkoxy radicals, which further attack intact unsaturated fatty acids, and the reaction proceeds in a chain manner. Thus, it is clear that iron complexes or iron ions are important for lipid peroxidation reactions in vivo. [0008] Furthermore, lactoferrin exists in intracellular and extracellular fluids such as milk and body fluids, and can take up two atoms of iron per molecule of protein, and has a strong affinity for iron. Lactoferrin, which is also synthesized within neutrophils and strongly captures iron ions, is known as a substance that inhibits bacterial growth and leads to death. Due to its iron affinity, iron, which is involved in lipid peroxidation reactions, is strongly incorporated into its protein molecules, so it is thought to act as a powerful antioxidant. Based on these matters, the present inventors conducted an investigation to investigate the relationship between lactoferrin and suppression of lipid peroxide production. As a result, it was confirmed that lactoferrin is an effective and safe lipid peroxide production inhibitor, and the present invention was completed. [0009] The present invention relates to a lipid peroxide production inhibitor containing lactoferrin as an active ingredient. [0010] The blending amount of the lipid peroxide production inhibitor containing lactoferrin as an active ingredient of the present invention is 0.00
Lactoferrin in a concentration of 1% by weight or more (hereinafter expressed in weight% unless otherwise specified), preferably 0.01% or more, is applied to the skin in various forms such as solution, powder, sol, gel, emulsion, wax, etc. Use by applying, rubbing, or spraying (aerosol). In addition, lactoferrin is blended into a known cosmetic or external preparation base by a conventional method.
lotions, skin creams, milky lotions, hair tonics,
It can be used as shampoos, conditioners, other cosmetics, pharmaceuticals, quasi-drugs (including various preparations for humans, other animals, and plants), and skin external preparations such as ointments. , lactoferrin is used as vitamins, animal and plant extracts, skin function enhancers, various surfactants, solvents,
It is stable against fragrances, colorants, bactericides, preservatives, antioxidants, humectants, other various additives, and various cosmetic bases, and can be blended with these and used in combination. [0011] The lactoferrin used in the present invention may be of any origin as long as it has the ability to bind iron, and lactoferrin obtained by tissue culture or chemical synthesis may also be used. Furthermore, as the method for producing lactoferrin, any known production method may be used. [0012] Below, in order to more specifically illustrate the present invention, the effects (action) or side effects (safety) of the lipid peroxide production inhibitor containing lactoferrin as an active ingredient will be explained.
It also discloses production examples, prescription examples, etc. of lactoferrin. [Production Example] Method for producing lactoferrin The ammonium sulfate precipitate of fresh beef whey is dissolved in water and desalted by passing it through a Sephadex G-25 column. The desalted protein was dissolved in phosphate buffered saline (PBS) at pH 7.3, and an anti-bovine lactoferrin monoclonal antibody was passed through an affinity column, followed by washing with PBS. Thereafter, bovine lactoferrin was eluted from the column with a 0.25M sodium acetate buffer with a pH of 3.7 containing 0.15M salt, and after adjusting the pH to around neutrality, it was dialyzed against purified water for 3 days, and then, Lyophilize the dialysate to obtain bovine lactoferrin. [Test Example 1] Confirmation test for the lipid peroxidation inhibitory effect of lactoferrin To determine whether lactoferrin inhibits the peroxidation of linolenic acid caused by ultraviolet rays, the amount of peroxide was measured using the TBA method. 1) Preparation of specimen Lactoferrin produced according to the manufacturing example or a known method is dissolved in purified water to prepare a specimen. In addition, as a control, vitamin E was tested in the same manner. 2) Test method 0.1% linolenic acid, 8 μM ferrous ammonium sulfate, and 38 μM ascorbic acid were added and dissolved in a 0.8% sodium lauryl sulfate aqueous solution, and 3.9 ml of this solution was dissolved in 10 ml.
Transfer to a screw bottle. Add 0.1 ml of the sample solution to this, and add ultraviolet light (Toshiba FL-20SE lamp and F
Three L-20SBLB lamps were arranged in parallel and irradiated at a distance of 15 cm. ) was irradiated for 1 hour, this solution was irradiated for 1 hour.
ml, then 0.8% thiobarbituric acid aqueous solution 1
．． Add 5 ml and 1.5 ml of 20% acetic acid (pH 3.5), and heat at 95°C for 1 hour. After cooling, 1 ml of purified water and 5 ml of n-butanol:pyridine (15:1) were added, shaken well, and centrifuged. The absorbance of this n-butanol layer at 532 nm was measured and determined as the amount of lipid peroxide. . In addition, the amount of lipid peroxide when adding a sample and irradiating ultraviolet rays is a, the amount of lipid peroxide when adding a sample and not irradiating ultraviolet rays, and the amount of lipid peroxide when irradiating ultraviolet rays without adding a sample. The lipid peroxide suppression rate was determined by equation 1, where a' was the amount of oxidized lipid, and b' was the amount of lipid peroxide when no sample was added and no ultraviolet rays were irradiated. 3) Test results As shown in Table 1, lactoferrin had a stronger inhibitory effect on lipid peroxidation than vitamin E. Also, from Table 1, the effective concentration range of lactoferrin is 0 in the prescription.
．． It was confirmed that a content of 0.001% or more is good. [Test Example 2] Safety test of lactoferrin [0021] 1) Preparation of specimen Using lactoferrin produced according to the manufacturing example or a known method, a soft lotion was prepared according to the following formulation and used as a specimen. As a control, purified water was added instead of lactoferrin. (Formulation) Ethanol
10.
0% Polyoxyethylene sorbitan monolaurate 1.0%
Methyl paraoxybenzoate

0.2% fragrance

0.2%
purified water

77.5% glycerin
5.0%
1,3-butylene glycol
6
．． 0% lactoferrin

0.1% [
2) Test method 30 healthy adults (11 men: 24-40 years old, 19 women)
A patch test was conducted on subjects (~46 years old) as controls. An appropriate amount of the sample was dropped onto a skin sensitization test tape: Fin Chamber (imported and sold by Taisho Pharmaceutical Co., Ltd.), and the tape was applied to the upper back for 47 hours. Judgment was made by determining skin erythema 1 hour and 24 hours after removal. 3) Test Results As shown in Table 2, it was determined that lactoferrin is a substance that can be safely used as a cosmetic raw material without any side effects. [0026] The method of the present invention will be further explained in detail by showing a few examples below. However, the field of application of the present invention is not limited to the following examples, but can be applied to various cosmetics, pharmaceuticals, etc. ,
It can be used in combination with quasi-drugs, etc. In addition, each example was manufactured by a conventional method for manufacturing each product, and
The lactoferrin used was the one produced in the above production example or a known method. [Example 1] (Formulation example of peel-off pack) Purified water
66.5% glycerin
5.0% propylene glycol 4.0
% polyvinyl alcohol
15.0% ethanol
8.0% polyoxyethylene glycol 1.0% methyl paraoxybenzoate 0.2% fragrance

0.2% lactoferrin
A peel-off pack containing 0.1% of the above formulation was manufactured. [Example 2] (Formulation example of cold cream) White beeswax
11.0% liquid paraffin
22.0% lanolin
10.0% almond oil
15.0% borax
0.5% methyl paraoxybenzoate 0.2% purified water
40.9% fragrance
0.3% lactoferrin
0.1% A cold cream was produced with the above formulation. [Example 3] (Example of shampoo formulation) Triethanolamine lauryl sulfate
5.0%
Polyoxyethylene lauryl ether sodium sulfate 12.0% 1,
3-butylene glycol
4.0%
Lauric acid diethanolamide
2.0%
Edetate disodium
0
．． 1% Methyl paraoxybenzoate
0.2% fragrance

0.3%
purified water

76.3%
lactoferrin
0.1%
A shampoo was manufactured using the above formulation. Effects of the Invention The effects of the lipid peroxide production inhibitor containing lactophilin as an active ingredient of the present invention can be summarized as follows.
It is as follows. ■ Significantly suppresses the production of lipid peroxide. ■It has no side effects and is extremely safe. Therefore, it can be used in various pharmaceuticals, quasi-drugs, cosmetics, and various foods (processed foods, functional foods, health foods, etc.).

Claims (1)

[Claims] 1. A lipid peroxide production inhibitor characterized by containing lactoferrin as an active ingredient.