KR102000550B1 - Composition comprising Polyamine compounds isolated from Quercus Mongolica pollen extracts for whitening - Google Patents

Abstract

The present invention provides a whitening composition comprising a polyamine compound selected from at least one of Mongolicine A (compound 1) and Mongoline A (compound 2), and more specifically, Relates to a whitening composition containing a polyamine-based compound separated from a Quercus Mongolica flowerpot.
The whitening composition is excellent in the activity of inhibiting tyrosinase activity and can be easily used in various forms of pharmaceutical composition, external preparation for skin, cosmetic product, and the like.

Description

[0001] The present invention relates to a whitening composition comprising a polyamine-based compound separated from a quince tree pollen (quercus mongolica pollen extracts for whitening)

The present invention relates to a whitening composition comprising at least one polyamine compound selected from the group consisting of Mongolicine A (Compound 1) and Mongoline A (Compound 2) isolated from Quercus Mongolica pollen .

Beauty has been a major concern of human beings since ancient times, according to the instinct of human pursuing beauty. Since modern life, the life expectancy of mankind has increased and the quality of life has become more important. One of the common elements of beauty, especially in the east and the west, is whiteness, and whitening is defined as whitening.

The history of whitening in Korea starts from the Chosun Dynasty. In Dangun Myth, it is described how to whitewash washed water after bathing with mugwort water or mixing garlic with honey. In the Three Kingdoms period, talc, rice flour, It is described how to make and apply whitening ingredients such as yellow loess, and how to apply skin care agent and whitening agent, which is a whitening agent, in Goryeo Dynasty. In the Joseon Dynasty, the distribution of hundreds of stores became active due to the sales of stores or waterworks. After the opening of the port, cosmetics such as cream and white powder were imported from the Qing Dynasty, and soft drinks (whitening lotion) and milky lotion It is said that it started. The history of whitening has been very long and, even in modern times, whitening among people's beauty activities is taking up a high proportion, and whitening methods that have been improved more than ever before are being researched in connection with the development of science and technology.

In the human body, skin acts as a protective membrane that protects living organisms from the external environment. It prevents the loss of biological components in the human body, such as water and electrolytes, and also protects harmful substances from entering the human body from the outside do. The skin is divided into the epidermal layer, the dermal layer and the subcutaneous fat layer, and the epidermal layer is composed of keratinocyte layer, granular layer, superficial layer, and basal layer. Melanocyte is formed in the base layer by making melanin. Melanin is a proteinaceous organic pigment that is formed from amino acids. It is widely distributed in living organisms. It plays a role in protecting skin from ultraviolet rays and maintaining body temperature.

Melanin is a factor that regulates hair color, eyes color and skin color of animals. It is classified into black and brown type of Eumelanin and yellow and red type of Pheomelanin. Melanin is synthesized by Tyrosinase ), TRP-1 (Tyrosinase related protein-1), and TRP-2 (Tyrosinase related protein-2). Tyrosinase, an enzyme that greatly affects the expression of melanin, is a rate-limiting enzyme for melanin biosynthesis. L-tyrosine is used as L-DOPA (3,4-dihydroxyphenylalanine) and L-DOPA It is converted into dopaquinone (DOPAquinone) and biosynthesizes melanin.

Melanin protects the skin from ultraviolet rays. However, excessive production of melanin causes pigmentation on the skin to form stains, freckles, dots, and black spots. If the intensity of ultraviolet light exposure is severe, it can cause skin cancer. Especially, modern people are exposed to ultraviolet rays more than before due to destruction of ozone layer due to environmental pollution. Therefore, the whitening of modern people is going beyond the aesthetic dimension and also on the health level.

The most popular whitening method is to apply cosmetics with added whitening ingredients. Whitening cosmetics can be broadly categorized into brightening or tone up and whitening products, The emphasis is on brightening the skin tone throughout the face, whitening products are focused on improving local pigmentation of the skin, such as stains and dullness. In the domestic whitening cosmetics market, whitening products are mainly produced.

Various ingredients such as Niacinamide, Alpha-Bisabolol, Ascorbyl Glucoside and Albutin have been continuously developed as whitening ingredients added to the whitening cosmetics. A brief review of the whitening mechanism of each ingredient shows that niacinamide inhibits melanin from migrating into the stratum corneum, while alpha-bisabolol inhibits melanin synthesis. Ascorbyl glucoside also inhibits the oxidation of tyrosine stimulated by tyrosinase enzyme, and arbutin inhibits the production of tyrosinase.

However, as many clinical studies on the risk of whitening ingredients have been made, the safety of whitening cosmetics continues to be controversial. For example, some customers using whitening cosmetics manufactured by famous cosmetics companies in Japan have suffered from side effects such as vitiligo, and the whitening cosmetics have been collected at 450,000. The cosmetics company has developed itself and has been certified by the Japanese Ministry of Health and Welfare, and Rhododenol, a compound that has the effect of inhibiting the activity of tyrosinase, is said to cause problems.

As another example, there is a case in which mercury in domestic whitening cosmetics exceeding the domestic limit value (ppm) by more than 15,000 times is detected. Mercury has a strong effect of blocking melanin production and has been widely used as a whitening cosmetic raw material in the past, but it has now been proven to cause nervous system disorders and its use is prohibited.

Arbutin, which is widely used in whitening cosmetics, is a compound in which hydroquinone and glucose are combined, but the binding between hydroquinone and glucose is easily decomposed at high temperatures, ultraviolet rays, enzymes, fermented products, Quinones and glucose are the most common cases. Hydroquinone has been shown to cause side effects such as vitiligo, skin allergies, contact dermatitis, and acne, and has been identified as a carcinogenic substance.

Kojic acid, which is widely used as a whitening cosmetic raw material together with arbutin, is inferior in in vivo inhibitory effect on melanin formation as compared with the in vitro inhibiting ability of tyrosinase, Recently, liver cancer has been reported to be caused by long-term use.

As the risk of such a whitening ingredient has been revealed, there are eight kinds of raw materials for whitening cosmetics at the phytosanitary clinic. Accordingly, safety whitening effect is excellent, and safety whitening composition free from side effects is desperately required. to be.

Pollen is a kind of granule-shaped germ cells contained in an anther in the operation of a plant (flower) and a flower (flower), and is formed in the operation of a flower. The shape of the pollen has a different shape depending on the type of plant and is piled up in a thick shell. There are many micropores in the shell, and there are germination holes that open the pollen tube. When the pollen is attached to the pistil, the pollen tube is extended through the germination hole, and the nucleus in the pollen tube is divided and divided into the support nucleus and the pollen tube nucleus. The pollen does not approach the pistil itself, so it is moved to the pistil by the force of the wind or insect and the correction is made. Bees produce propolis, honey, royal jelly, beeswax, bee pollen and have been used medicinally in various countries including Egypt, Greece, and China.3) -7) Recently, , Are effective as anticancer drugs and have recently been used as an alternative drug without side effects. The reptiles are made up of dozens to millions of pollen grains, which move like dumplings using their saliva on their legs to provide nutrients to young bees. Pollen is a protein source of larvae and adults of bees and is richer in nutrients than ordinary pollen. It has been widely used in many countries including Korea for edible or medicinal purposes. It is known to be rich in carbohydrates, fats, vitamins, minerals, and has excellent physiological functions. The main constituents of the pollen are protein (5-60%), essential amino acid, reducing sugar (13-55%), fat (4-7%), nucleic acid and fiber (0.3-20% There are various nutrients to be clear. Other minerals such as Ca, Mg, Fe, Zn, and Cu are also abundant. These include pro-vitamin A (β-carotene), vitamin E (tocopherol), niacin, thiamin, biotin, . Among physiologically active substances, unsaturated fatty acid, saturated fatty acid, phospholipid (1.5%), plant sterol, β-sitosterol, p-sitosterol (1.1%) and terpene are known. Polyphenol and flavonoid (3-8%) are known. Phenolic acids and flavonoids have been reported as typical polyphenol compounds in pollen. Phenolic aicd has been reported to contain chlorogenic acid, gallic acid, ferulic acid, cinnamic acid, cinnamic acid, hydroxycinnamic acid and caffeic acid. The compounds are luteolin, apigenin, chrysin, quercetin, rutin, kaempherol, myricetin, galangin, naringenin, pinocembrin, genistein and isorhamnetin. It is known that aristotle, hippocrates, pythagoras, pliny, etc. of the ancient medieval western medical literature are very effective for the soothing, gastrointestinal and cardiovascular diseases. Pollen is known to have high physiological effects such as nutritional effects, dressing, neuropathy, arteriosclerosis and geriatric diseases. Pollen has been reported to be effective in the circulatory system, digestive system, metabolic system, anti-aging, etc. Recent studies have shown that antioxidant, antiinflammatory, anticarcinogenic, antibacterial, antifungal, antioxidant, immune enhancement, atherosclerosis, .

Quercus mongolica ( Quercus mongolica ) is a member of the Quercus family (Fagaceae). It is widely distributed in Korea, Japan and China, and domestically occupies most of the natural forests in Korea. Fruits are picked in autumn and dried in the sun for food use. Oakwood is used for timber, building materials, cork materials, and equipment. In the private sector, stem and skin were dried and used for atopy, used for gastrointestinal disorders, cough, and used as a remedy for hemostatic agents and diarrhea in Oriental medicine. The physiological activity has been reported to be effective for antitumor, antiviral, antiallergic and antioxidant activities. The leaves and bark of Quercus mongolica contain flavonoids and tannins. They have been used for oral inflammation with excellent efficacy against cerebral hemorrhage and jaundice. In addition, a wide variety of flavonoids and tannins of polyphenols have been reported as active ingredients of Quercus mongolica, including ellagic acid, catechin, taxifolin, glucodistylin, Mongolicanin, mongolinin, and acutissimin C have been reported to be involved.

The inventors of the present invention found that the polyamine compound isolated from the extract of Quercus mongolica can be used as a whitening pharmaceutical composition or a cosmetic composition because of its excellent whitening effect and no skin side effect while studying the efficacy of the Quercus mongolica .

Korean Registered Patent No. 10-1483872 (Name of the invention: composition for anti-wrinkle including fractions of Quercus mongolica leaf, Applicant: Chung-Ang University Industry-Academic Collaboration Foundation, Registered Date: 2015.01.12) Korean Patent No. 10-1224238 (name of the invention: mixed plant extract, applicant: Jung Soo Kee, publication date: 2013.01.14) Korean Patent No. 10-0992087, entitled "Cosmetic Composition Containing Quercus mongolica Leaf, Shulp, and Radish Extract Mixture, Applicant: Kosemeca Korea Co., Ltd., Registered on Oct. 29, 2010)

Almaraz-Abaraca, N .; da Graca Campos, M .; . G. C., M .; Avila-Reyes, J. A .; Naranjo-Jimenez, N .; Herrera-Corral, J .; Gonzales-Valdez, L. S. Variability of antioxidant activity among honeybee-collected pollen of different botanical origin. Interciencia-Caracas 2004, 29, 574-578.

It is an object of the present invention to provide a whitening agent containing at least one polyamine compound selected from Mongolicine A (Compound 1) and Mongoline A (Compound 2) isolated from Quercus Mongolica pollen To provide a composition.

The present invention relates to a whitening composition comprising at least one polyamine compound selected from the group consisting of Mongolicine A (Compound 1) and Mongoline A (Compound 2) .

[Chemical Formula 1]

(2)

The compound is distinguished from quercus mongolica pollen.

The compound is characterized by inhibiting the activity of tyrosinase.

The present invention also relates to a cosmetic composition for whitening characterized by containing a polyamine compound selected from at least one of Mongolian lysine A (Compound 1) and Mongolin A (Compound 2).

Accordingly, the present invention can provide a cosmetic composition containing the cosmetic composition as a raw material or an effective ingredient.

The present invention also provides a method for separating said polyamine-based compound from quinoa pollen.

Preferably, the method comprises the steps of extracting Quercus mongolica pollen with alcohol or an aqueous solution thereof to prepare Quercus mongolica pollen extract; Sequentially fractionating the Quercus mongolica extract with hexane, dichloromethane (CH ₂ Cl ₂ ), ethylaldehyde (EtOAc) and n-butanol (BuOH) to prepare respective fractions; And fractionating the n-butanol fraction of the fraction, wherein the fraction is selected from the group consisting of Mongolian lysine A (Compound 1) and Mongol Line A (Compound 2).

The present invention relates to a novel compound of Mongolicine A (Compound 1) of Formula 1 or Mongoline A (Compound 2) of Formula 2 separated from Quercus mongolica.

Hereinafter, the present invention will be described in more detail.

The present invention relates to a whitening composition comprising a polyamine-based compound selected from at least one of Mongolian lysine A (Compound 1) and Mongolian Line A (Compound 2), said compound being preferably selected from the group consisting of alcohols Extracting the extract with an aqueous solution thereof to prepare a Quercus variabilis extract; Sequentially fractionating the Quercus mongolica extract with hexane, dichloromethane (CH ₂ Cl ₂ ), ethylaldehyde (EtOAc) and n-butanol (BuOH) to prepare respective fractions; And performing the chromatography in the n-butanol fraction of the fraction.

The alcohol may be a C1-C4 alcohol or a mixed solution thereof, and the C1-C4 alcohol may be selected from the group consisting of methanol, ethanol, propanol, isopropanol, butanol and isobutanol.

The alcohol may be preferably ethanol or methanol, and the aqueous solution of alcohol may be 70-90% (v / v) aqueous solution thereof. The alcohol extraction may be performed twice or more, but is not limited thereto. The alcohol extraction may be performed twice or more, but is not limited thereto.

The compound may be isolated from various parts such as leaves, flowers, branches of Quercus mongolica, but is not limited thereto. Most preferably from the pollen.

The polyamine-based compound may be obtained by preparing a quinceanera pollen extract or a fraction thereof, followed by filtration, concentration, fraction extraction, separation, and the like.

The weight ratio of the quince seed pollen sample and the alcohol may be variously set when preparing the Quercus mongolica pollen extract using alcohol, and most preferably, 300 to 1000 parts by weight of the alcohol may be set based on 100 weight parts of the Quercus mongolica sample But is not limited thereto.

The extracts or fractions of Quercus mongolica pollen may be separated by chromatography. The chromatography may be carried out using silica gel column chromatography, LH-20 column chromatography, ion exchange resin chromatography (Eg, ion exchange resin chromatography, medium pressure liquid chromatography, thin layer chromatography (TLC), silica gel vacuum liquid chromatography, size- exclusion chromatography, preparative chromatography, and high performance liquid chromatography. Preferably, high performance liquid chromatography can be selected, but is not limited thereto.

The elution solvent used in the selection of the high performance liquid chromatography may be variously selected, and preferably water, methanol or a mixture thereof may be selected, but is not limited thereto.

In addition, the mixing ratio of the solvent may be variously selected, and most preferably, it is not limited to 75% (volume) of methanol and 25% (volume) of water.

The fractions of the Quercus mongolica extract thus isolated can be further fractionated. Additional fractionation can be selected among the chromatographies listed above.

Preferably, medium-pressure liquid chromatography can be selected, but is not limited thereto. When the medium pressure liquid chromatography is selected, a variety of elution solvents may be selected, and dichloromethane, methanol or a mixture thereof may be preferably selected, but is not limited thereto. Also, the mixing ratio of the solvent may be variously selected, and most preferably, it may be 75% by volume of dichloromethane and 25% by volume of water, but is not limited thereto. LH-20 column chromatography can be preferably selected, but is not limited thereto. When LH-20 column chromatography is selected, a variety of solvents may be selected, and methanol is preferably used, but not limited thereto. In addition, chromatography for preparations may be selected, but not limited thereto. When the chromatography for production is selected, the solvent may be variously selected, and preferably water, methanol or a mixture thereof may be selected, but is not limited thereto. Also, the mixing ratio of the solvent may be variously selected, and most preferable is 50 to 55 (vol.)% Of methanol and 45 to 50 (vol.%) Of water.

The equipment to be used in the extraction, filtration, concentration, fraction extraction, separation and the like of the present invention may be any of those conventionally used, and the extract or fraction and the compound to be finally isolated may be subjected to various methods such as hot air drying, To remove the solvent.

The polyamine compound selected from at least one of Mongolian Lysine A (Compound 1) and Mongolian Line A (Compound 2) of the present invention has an effect of inhibiting the activity of tyrosinase and can be usefully used for skin whitening.

Accordingly, the present invention can provide a whitening pharmaceutical composition or a skin external preparation containing a polyamine compound selected from the group consisting of Mongolian lysine A (compound 1) and Mongol line A (compound 2) and a pharmaceutical excipient.

At this time, the pharmaceutical composition or the external preparation for skin may be administered to mammals such as rats, livestock, and humans in various routes. The pharmaceutical composition may be administered orally or vascularly. Preferably, the pharmaceutical composition may be administered orally or parenterally such as a spray, a liquid preparation, a lotion, a foaming agent, a tape agent, an aerosol agent, an external preparation, a gel, a cream, , A patch, a mask pack or the like. The ingredients other than the active ingredient of the pharmaceutical composition or external preparation for skin of the present invention may be any ingredient commonly used in external preparations applied to the skin. For example, bases such as white petrolatum (vaseline), yellow petrolatum, lanolin, purified beeswax, cetanol, stearyl alcohol, stearic acid, hydrogenated oil, hydrocarbon gel, polyethylene glycol, liquid paraffin and squalane; Oleic acid, isopropyl myristate, glycerol triisooctanoate, crotamitone, diethyl sebacate, diisopropyl sebacate, diisopropyl adipate, hexyllaurate, fatty acid, fatty acid ester, vegetable oil, Solvents or stabilizers such as alcohols; Tocopherol derivatives, antioxidants such as L-ascorbic acid, dibutylhydroxytoluene and butylhydroxyanisole; preservatives such as p-hydroxybenzoate; Curling agents such as glycerin, flufolene glycol and sodium hyaluronate; Surfactants such as polyoxyethylene derivatives, glycerol fatty acid esters, sucrose fatty acid esters, sorbitan fatty acid esters, propylene glycol fatty acid esters and lecithin; Thickeners such as carboxyvinyl polymer, xanthan gum, carboxymethylcellulose and sodium carboxymethylcellulose, hydroxypropylcellulose and hydroxypropylmethylcellulose; Stabilizers; Preservatives; Adsorption promoters; And other suitable fillers may be added. The pharmaceutical composition or external preparation for skin of the present invention may be prepared by mixing well the components well known in the art for preparing external preparations, together with a suitable base, if necessary, and the preparation thus prepared is applied to lesions as required .

The dosage of the pharmaceutical composition or external preparation for skin of the present invention will vary depending on the age, sex, body weight of the subject to be treated, and the severity of the pathological condition, the disease or pathological condition, the route of administration and the judgment of the prescriber. Dosage determinations based on these factors are within the level of ordinary skill in the art and generally the dosage ranges from 0.01 mg / kg / day to approximately 2000 mg / kg / day. A more preferable dosage is 1 mg / kg / day to 500 mg / kg / day. The administration may be carried out once a day or several times. The dose is not intended to limit the scope of the invention in any way.

The present invention also relates to a whitening cosmetic composition containing a polyamine compound selected from at least one of Mongolian lysine A (Compound 1) and Mongol Line A (Compound 2). Accordingly, the present invention can provide a cosmetic composition containing the cosmetic composition as a raw material or an effective ingredient. The formulations of the cosmetics may be prepared in any cosmetic formulations conventionally produced in the art and may be formulated into ointments, essences, whitening creams, lotions, emulsions, packs, general lotions, skin milks, skins, creams, Soap, softening lotion, medicated lotion, whole body cleanser, cleansing oil, cleansing cream, cleansing tissue and cleansing water. The compound of the present invention contained in the cosmetic preparation is 0.0001 to 10.0% by weight, preferably 0.0005 to 10.0% by weight, and most preferably 0.005 to 5.0% by weight based on the total cosmetics. If the amount is less than 0.0001% by weight, a pronounced whitening effect can not be expected, and it may be more than 10.0% by weight, but it is not preferable from the viewpoint of cost.

The cosmetic preparation of the present invention may contain an ingredient selected from the group consisting of animal oils, vegetable oils, waxes, paraffin, starch, tragacanth, cellulose derivatives, polyethylene glycol, silicone, bentonite, silica, talc, zinc oxide, lactose, But are not limited to, calcium silicate, polyamide powder, chlorofluorohydrocarbons, propane / butane or dimethyl ether, water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propyleneglycol, Glycerol aliphatic esters, polyethylene glycols or sorbitan fatty acid esters, ethoxylated isostearyl alcohols, polyoxyethylene sorbitol esters and polyoxyethylene sorbitan esters, microcrystalline cellulose, aluminum metahydroxide, bentonite, Aga or Trakant, Province But are not limited to, alcohol sulfates, aliphatic alcohol ether sulfates, sulfosuccinic acid monoesters, isethionates, imidazolinium derivatives, methyltaurate, sarcosinate, fatty acid amide ether sulfates, alkylamidobetaines, fatty acid glycerides, fatty acid diethanolamides, Lanolin derivatives or ethoxylated glycerol fatty acid esters.

The present invention also provides a whitening health functional food containing a polyamine compound selected from at least one of Mongolian lysine A (Compound 1) and Mongol Line A (Compound 2). The polyamine compound selected from the group consisting of Mongolian lysine A (compound 1) and Mongol line A (compound 2) may be added in an amount of 0.001 to 30% by weight to the health functional food of the present invention. The health functional food of the present invention includes forms such as tablets, capsules, pills, and liquids. Examples of the foods to which the extract of the present invention can be added include various kinds of drinks, meat, sausage, bread, Snacks, noodles, ice cream, dairy products, soups, ionic drinks, beverages, alcoholic beverages, gums, tea and vitamin complexes.

The present invention relates to a whitening composition comprising a polyamine-based compound selected from at least one of Mongolian lysine A (Compound 1) and Mongolian Line A (Compound 2) isolated from Quercus Mongolica pollen. The whitening composition is excellent in inhibiting activity against tyrosinase and can be easily used in various forms of pharmaceutical composition, external preparation for skin and cosmetic product.

Korean Patent No. 10-101483872, Korean Patent No. 10-1224238, and Korean Patent No. 10-0992087 disclose a cosmetic composition containing the extract of Quercus mongolica and the like, The whitening composition prepared by containing the compound has not been disclosed at all.

Figure 1 shows the process for the preparation of extracts and fractions of Quercus mongolica pollen.
Fig. 2 shows the step of re-fractionating the n-butanol fraction from the fractions of the Quercus mongolica pollen.
Fig. 3 shows a step of further fractionating B75M fraction of Quercus mongolica pollen extract.
Figure 4 shows the HRESI-TOF-MS spectrum results of Mongolian lysine A (Compound 1).
Figure 5 shows the HMBC spectrum correlation of Mongolian lysine A (Compound 1).
6 shows the HRESI-TOF-MS spectrum results of Mongolia line A (compound 2).
Figure 7 shows the HMBC spectrum correlation of Mongolia line A (compound 2).

Hereinafter, preferred embodiments of the present invention will be described in detail. However, the present invention is not limited to the embodiments described herein but may be embodied in other forms. Rather, these embodiments are provided so that this disclosure will be thorough and complete, and will fully convey the concept of the invention to those skilled in the art.

≪ Example 1: Isolation of polyamine-based compound >

Quercus mongolia pollen samples were used in Rural Development Administration in February 2015, and reagents and solvents were used for analytical grade or grade 1 reagents. HPLC grade solvents were used for HPLC.

15.0 kg of Quercus mongolica pollen was extracted twice with 90 L of 80% methanol at room temperature. The extract was filtered with a vacuum filter, filtered and the filtrate was concentrated with a rotary evaporator to obtain an extract. The residue was suspended in distilled water and fractionated in the order of n-hexane, dichloromethane (CH ₂ Cl ₂ ), ethyl acetate (EtOAc), n-butanol (BuOH) A dichloromethane fraction, an ethyl acetate fraction, an n-butanol fraction and a water fraction were obtained (see Fig. 1).

Among them, the n-butanol fraction was suspended in distilled water and subjected to HP-20 column chromatography under the conditions of adjusting the specific gravity of water and methanol to 0%, 25%, 75%, and 100%, respectively. Fractions of B50M, B75M and B100M were obtained (see Fig. 2).

The B75M fractions were subjected to MPLC silica column chromatography under the conditions of adjusting the specific gravity of dichloromethane and methanol from 100% to 0%, respectively. Twelve small fractions (B75M-1 to B75M-12) (B75M-3-1 to B75M-3-11) were obtained by performing Sephadex LH-20 column chromatography under the conditions of 100% methanol elution, .

The B75M-3-3 fraction of the 11 small fractions was subjected to preparative HPLC under the elution conditions of methanol 55 (volume)% and water 45 (volume)%, and Compound 1 was isolated. 4 fractions (B75M-4-1 to B75M-4-4) were obtained by performing Sephadex LH-20 column chromatography under the elution condition of 100% methanol. The B75M-4-1 fraction was subjected to preparative HPLC under the elution conditions of 50% by volume of methanol and 50% by volume of water to separate the compound 2.

<Example 2> Identification of physicochemical structure of polyamine-based compound derived from Quercus mongolica tree>

Example 2-1. Mongolicine A (compound 1)

The physicochemical properties and spectroscopic data of Mongolian lysine A (Compound 1) are as follows.

White amorphous powder;

: -64.2° (c 0.017, MeOH); [alpha]

: -64.2 [deg.] (C 0.017, MeOH);

FT-IR? _Max 3327, 1597, 1229 cm ^-1 ;

UV (MeOH) [lambda] _max 289 (1.81) nm;

HRESI-TOF-MS m / z 387.1889 [M + Li] + (calcd. For C 22 H 2 4N 2 O 4 387.1896).

Table 1 below shows the results of ¹ H-NMR (CD ₃ OD, 700 MHz) and ¹³ C-NMR (CD ₃ OD, 175 MHz) of Mongolian lysine A (Compound 1).

Mongolian lysine A was isolated and purified as a white amorphous powder and its molecular weight was confirmed by HRESI-TOF-MS spectrum at m / z 387.1889 (calcd. For C ₂₂ H ₂ 4N ₂ O ₄ 387.1896). The structure of Mongolian lysine A is similar to that of di- p- coumaroylputrescine (Interciencia-Caracas 2004, 29, 574-578), which is a known compound. The trans-double bond peak δ _H 6.42 (1H, d, J = 16.1 Hz, H-2 '), δ C 117.0 (C-2'), δ H 7.46 (1H, d, J = 16.1 Hz, H-3 '), δ C 140.4 (C-3') and cis- double bond _{peak δ H 5.85 (1H, d} , J = 12.6 Hz, H-2 "), δ C 120.3 (C-2"), δ H 6.65 (1H, d, J = 12.6Hz, H-3 was confirmed _{"), δ C 136.5 (C} -3").

And two 1,4-substitued aromatic corresponding to the _{ring δ H 7.42 (2H, d} , J = 9.1 Hz, H-5 ', 9'), δ C 129.1 (C-5 ', 9'), δ H 6.81 (2H, d, J = 9.1 Hz, H-6 ', 8'), δ C 115.3 (C-6 ', 8'), δ C 126.3 (C-4 '), δ C 159.1 C-7' ) peak and _{δ H 7.41 (2H, d,} J = 9.1 Hz, H-5 ", 9"), δ C 130.8 (C-5 ", 9"), δ H 6.75 (2H, d, J = 9.1 Hz , H-6 ", 8"), δ _C 114.6 (C-6 ", 8"), δ _C 126.7 (C-4 ") and δ _C 157.8 (C-7"

The molecular formula C ₂₂ H ₂ 4 N ₂ O ₄ was predicted through the above HRESI-TOF-MS spectrum (see FIG. 4) and ¹ D NMR data.

H-2 '/ C-4', H-3 '/ C-1', 5 ', 9' and H-2 "/ C- C-1 and H-6 / C-1 ", confirming the presence of trans- and cis- coumaroyl groups by confirming the correlation between" 6 , it was confirmed that one trans-, cis- coumaroyl group was substituted for each (see Fig. 5).

Based on the above analysis results of Compound 1 is ^{N 1 - (E) - N} 6 - (Z) - di - p - Koh mellow days Trail New Fu ^{(N 1 - (E) -} N 6 - (Z) -di- p -coumaroyl putrescine). This compound was first reported in nature and was named Mongolicin A (Mongolicin A).

Example 2-2. Mongoline A (Compound 2)

The physicochemical properties and spectroscopic data of Mongolia line A are as follows.

White amorphous powder;

: -107.2° (c 0.007, MeOH);[alpha]

: -107.2 DEG (c 0.007, MeOH);

FT-IR? _Max 3369, 1601, 1251 cm ^-1 ;

UV (MeOH) [lambda] _max 280 (1.74);

HRESI-TOF-MS m / z 793.3785 [M + Li] ⁺ (calcd. For C ₄₆ H ₅₀ N ₄ O ₈ 793.3789).

Table 2 below shows the results of ¹ H-NMR (CD ₃ OD, 700 MHz) and ¹³ C-NMR (CD ₃ OD, 175 MHz) of Mongolia line A (compound 2).

Mongolia line A was isolated and purified as white amorphous powder and its molecular weight was confirmed by HRESI-TOF-MS m / z 793.3785 [M + Li] ⁺ (calcd. For C ₄₆ H ₅₀ N ₄ O ₈ 793.3789).

^{The 1} H-NMR and ¹³ C-NMR spectra showed overlapping resonances and the four cis- double bonds δ _H 5.74 / 5.75 (2H, d, J = 12.6 Hz, H-2 ' _{) δ H 5.78 / 5.79 (2H} , d, J = 12.6 Hz, H-2 ', 2 ""), δ C 121.2 (C-2', 2 ""), δ H 6.50 (1H, m, H- 3 ', 3 ""), δ C 136.8 / 137.0 (C-3', 3 ""), δ H 5.84 / 5.86 (2H, d, J = 12.8 Hz, H-2 ", 2"'), δ _{H 5.93 / 5.95 (2H, d} , J = 12.8 Hz, H-2 ", 2"'), δ C 121.4 (C-2 ", 2"'), δ H 6.39 / 6.46 (2H, d, J = 12.8 Hz, H-3 ", 3"') and δ _C 132.2 (C-3 ", 3"').

In addition, _{1,4-substitued aromatic ring δ H 7.61} (4H, d, J = 8.4 Hz, H-5 ', 9', 5 "", 9 ""), δ C 132.3 (C-5 ', 9', 5 "", 9 "") , δ H 6.71 (4H, d, J = 8.4 Hz, H-6 ', 8', 6 "", 8 ""), δ C 115.6 / 115.7 (C-6 ', 8 ', 6 "", 8 ""), δ C 126.8 (C-4', 4 ""), δ C 158.4 (C-7 ', 7 ""), δ H 7.19 (2H, dd, J = 8.4 Hz, 2.8, H-5 ", 5"'), δ H 7.24 (2H, t, J = 8.4 Hz, H-5 ", 5"'), δ C 130.4 (C-5 ", 5"' _{), δ H 6.70 (2H,} d, J = 8.4 Hz, H-6 ", 6"'), δ C 115.2 (H-6 ", 6"'), δ C 121.4 (C-4 ", 4"') and δ _C 158.1 / 158.2 (H-7 ", 7"').

The molecular formula C ₄₆ H ₅₀ N ₄ O ₈ was predicted by the above HRESI-TOF-MS spectrum (see FIG. 6) and ¹ D-NMR data.

H-2 '/ C-4', H-3 '/ C-1', 5 ', 9' 5 &quot;, 9 &quot;, H-2 "/ C-4 &quot;, H- H-3 "" / C- 1 "", 5 "", 9 " check the correlation between" was confirmed by a group of four cis- coumaroyl, H-2 / C -3, 4, H-4 / C- 3.0-3.5 ppm via H-6 / C-7, 8, H-9 / C-7, 8, H-11 / C-9, 12, 13, H- H-2 / C-1 ', H-4, 6 / C-1 ", H-9, and 11 / C-1""of the methylene peaks of 1.1-1.7 ppm were found to be spermine- , H-13 / C-1 "&quot;, confirming that the position of the cis-coumaroyl group was substituted at positions 1, 5, 10 and 13 (see FIG.

Based on the above analysis of the compound ^{2 N 1, N 5, N} 10, N 14 - (Z) - tetrahydro -p- Koh mellow one spermine ^{(N 1, N 5, N} 10, N 14 - (Z) - tetra-p-coumaroyl spermine), which was first reported in nature and was named Mongoline A (Mongoline A).

<Experimental Example 1: Effectiveness of tyrosinase inhibitory activity>

Add 1 μl of each of Compounds 1 and 2 isolated from Quercus mongolica pollen to a 96-well plate. Add 50 μl of a solution in which 1 mM of tyrosine, a substrate, is dissolved in PBS (phosphate buffered saline) And allowed to stand for 5 minutes.

Tyrosinase diluted at a concentration of 10 units / ml was diluted to a concentration of 0.1 unit / ml in PBS, and 50 ㎕ was added to each well. The plate was allowed to stand for 15 minutes. The absorbance was measured at a wavelength of 490 nm using a microplate reader. Kojic acid was used as a positive control. The results are shown in Table 3 by comparing IC ₅₀ values of Mongolian lysine A (compound 1), Mongolian line A (compound 2) and kojic acid Tyrosnase inhibitory activity.

Tyrosnase inhibitory activity IC ₅₀ values (μM) Mongol lysine A (Compound 1) 75.1 Mongolia line A (compound 2) 19.5 Kojic acid 36.4

Table 3. Note the Mongolian lysine A (Compound 1) in inhibitory activity IC ₅₀ is Tyrosnase to 75.1μM, Mongolian line A Tyrosnase inhibitory activity IC ₅₀ (Compound 2) is confirmed to 19.5μM excellent tyrosinase inhibition activity .

&Lt; Preparation of cosmetic preparation 1. Preparation of soap >

2 wt% of olive oil, 5 wt% of palm oil, 20 wt% of sorbitol, 0.2 wt% of carbomer, 0.5 wt% of hydrogenated castor oil, 10 wt% of dipropylene glycol, 0.15 wt% of phenoxyethanol, An emulsion comprising 0.02 wt.% Of acetic acid, 69.13 wt.% Of water, 1 wt.% Of Mongol lysine A (Compound 1) or Mongol Line A (Compound 2) of Example 2 and 0.2 wt.% Of triethanolamine was prepared. Soap was prepared by mixing 90 g of a natural soap base sold in the market.

&Lt; Preparation of cosmetic preparation 2. Preparation of skin >

1% by weight of Mongol lysine A (Compound 1) or Mongol Line A (Compound 2) of Example 2, 5.2% by weight of propylene glycol, 1.5% by weight of oleyl alcohol, 3.2% by weight of ethanol, 3.2% Skin was prepared using a conventional method with 2.0% by weight of phenone-9, 1.0% by weight of carboxyl vinyl polymer, 3.5% by weight of glycerin, a small amount of preservative, and a residual amount of purified water.

&Lt; Cosmetic preparation example 3. Lotion production >

1 wt% of Mongolian lysine A (Compound 1) or Mongol Line A (Compound 2) of Example 2, 1.0 wt% of cetostearyl alcohol, 0.8 wt% of glyceryl monostearate, 0.3 wt% of sorbitan monostearate, A mixture of 1.0 wt% of Solvite 60, 5.0 wt% of mineral oil, 3.0 wt% of cyclomethicone, 0.5 wt% of dimethicone, 0.1 wt% of allantoin, 5.0 wt% of glycerin, 2 wt% of alcohol, 3.0 wt% of propylene glycol, A lotion was prepared using a conventional method in terms of the amount of the antiseptic agent and the amount of the purified water.

&Lt; Cosmetic preparation example 4. Essence preparation >

1% by weight of propylene glycol, 3.0% by weight of glycerin, 0.5% by weight of allantoin, 0.01% by weight of EDTA-2Na and 5.0% by weight of ethanol were dissolved in 1% by weight of Mongolian lysine A (Compound 1) , 1.5 wt% of triethanolamine, 2.0 wt% of squalane, 2.5 wt% of beeswax, 60 wt% of polysorbate, 1.0 wt% of carboxyl vinyl polymer, 2.5 wt% of sorbitan sesquioleate, The contents were used to prepare essences using conventional methods.

&Lt; Preparation of cosmetic preparation 5. Cream production >

1% by weight of Mongol lysine A (Compound 1) or Mongol Line A (Compound 2) in Example 2 of the present invention, 0.7% by weight of polyoxyethylene sorbitan monostearate, 0.5% by weight of sorbitan sesquioleate, 0.6% by weight of cetyl alcohol 0.6 By the conventional method with respect to the content of stearic acid, 0.75 wt% of stearic acid, 0.6 wt% of glyceryl monostearate, 15.0 wt% of liquid paraffin, 10.0 wt% of carboxyvinyl polymer, 0.2 wt% of triethanolamine, To prepare a cream.

&Lt; Formulation Example 1 >

1 g of Mongol lysine A (Compound 1) or Mongol Line A (Compound 2) of Example 2 of the present invention, 0.6 g of sodium chloride and 0.1 g of ascorbic acid were dissolved in distilled water to make 100 ml. This solution was placed in a bottle and sterilized by heating at 20 DEG C for 30 minutes.

<Formulation Example 2: Food Preparation>

Formulation Example 2-1. Manufacture of cooking seasonings

An extract containing Mongolian lysine A (Compound 1) or Mongolian Line A (Compound 2) of Example 2 of the present invention was added to the cooking sauce in an amount of 1 wt% to prepare a cooking sauce for health promotion.

Formulation example  2-2. Manufacture of flour food products

An extract containing Mongolian Lysine A (Compound 1) or Mongolian Line A (Compound 2) of Example 2 of the present invention was added to the flour at 0.1 wt%, and the mixture was used to prepare breads, cakes, cookies, crackers and noodles To produce health promotion foods.

Claims (10)

delete delete delete delete delete delete delete Extracting and preparing the extract of the pollen Oak Wood Oak Wood (Quercus Mongolica) pots with an alcohol or an aqueous solution thereof;
Sequentially fractionating the Quercus mongolica extract with hexane, dichloromethane (CH ₂ Cl ₂ ), ethylaldehyde (EtOAc) and n-butanol (BuOH) to prepare respective fractions; And
Performing the chromatography in the n-butanol fraction of the fraction;
Wherein the polyamine compound is selected from the group consisting of compounds represented by the following formulas (1) and (2).
[Chemical Formula 1]

(2)

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