TW201946913A - Use of green mango extracts and compounds obtained therefrom for skincare and healthcare - Google Patents

Abstract

The present invention provides use of a green mango extract for manufacture of a pharmaceutical composition for UV protection and whitening of skin and anti-glycation. The present invention also provides compounds isolated from the green mango extract and identified as hydrolyzed tannins, and use of said compound for manufacture of a pharmaceutical composition for anti-glycation.

Description

   Mango young fruit extract and use thereof for skin care and health care   

The present invention relates to the use of a young mango fruit extract and the hydrolyzed tannin compound obtained therefrom, and particularly to the use of a young mango fruit extract to prepare skin anti-ultraviolet and whitening and anti-glycation pharmaceutical compositions, and Use of a hydrolyzed tannin compound for preparing an anti-glycation pharmaceutical composition.

Skin provides the first line of protection for the body against UV radiation from the sun. The skin includes the epidermis (including the outermost stratum corneum), the dermis, and subcutaneous tissue. The epidermis is the outermost layer of the skin and is constantly renewed. There are continuously dividing cells (such as fibroblasts, keratinocytes, and melanocytes) between the epidermis and the dermis. The activities of these cells are very sensitive to ultraviolet rays. The dermis contains collagen and elastin, which give the skin elasticity and support. When exposed to high amounts of ultraviolet light (mainly ultraviolet A), collagen fibers and elastin are damaged, resulting in reduced skin elasticity, wrinkle formation, and the appearance of skin aging. In order to delay skin aging, it is extremely important to reduce the damage to skin caused by ultraviolet radiation.

Fair and translucent complexion is also a symbol of youthful energy. In order to show personal vitality, modern people maintain skin whiteness through methods such as makeup and skin care. Skin tone is mainly determined by the amount of melanin produced by melanocytes. Congenital heredity, endocrine disorders, daily life, sun exposure, and drug use may all cause melanocytes to produce melanin, which may lead to dull skin or local spots. Therefore, the development of a method for actively inhibiting melanin production is the key to maintaining bright skin.

Another cause of skin aging is glycation, which is a non-enzymatic reaction between the aldehyde (ketone) group of a reducing sugar and the amine group of an amine-containing substance (eg, protein, nucleic acid, etc.). When the human body is in a state of hyperglycemia for a long time, proteins in the body are susceptible to glycation reactions with glucose, eventually forming advanced glycation end products (AGEs). In addition to the body's own synthesis, end-glycation products also enter the body through diet. Overcooked or processed foods often contain higher amounts of end-saccharification products, such as grilled foods, fries, bread, and the like. The protein that forms the end-glycation product will lose its normal molecular structure and physiological functions.At the same time, the end-glycation product will also increase the oxidative stress of the cells and induce the secretion of pro-inflammatory cytokines, so it may trigger a variety of metabolic diseases that cause the body to age, such as atheromatosis. Arteriosclerosis, neurodegenerative diseases, cataracts, kidney failure, etc. As far as skin aging is concerned, the glycation reaction will affect the aggregation of collagen to form fibers, and the glycated collagen fibers become stiff and brittle, making the skin prone to wrinkles. Therefore, inhibiting the occurrence of glycation reaction is also a countermeasure for slowing skin aging.

Currently, most of the sunscreen, whitening, or anti-glycation ingredients in commercially available skin care products are chemically synthesized. If these ingredients are not used properly, they may be harmful to skin health. Therefore, it is necessary to develop natural ingredients with skin whitening, anti-ultraviolet, and anti-glycation to prepare new skin care products.

One object of the present invention is to provide a young mango fruit extract for preparing skin anti-ultraviolet and whitening and anti-glycation pharmaceutical composition, wherein the young mango fruit extract is obtained by extracting a young mango fruit with a solvent, wherein The young mango fruit line is an immature mango fruit with a length of 3 to 7 cm.

In one embodiment of the present invention, the weight ratio of the solvent to the young mango fruit ranges from 20: 1 to 1: 1, and the extraction is performed at 55 ° C to 100 ° C.

In one embodiment of the present invention, the solvent is water, and the concentration of the young mango fruit extract is at least 0.25 mg / mL.

In one embodiment of the present invention, the mango young fruit extract inhibits melanin formation, prevents and repairs damage to skin fibroblasts caused by ultraviolet A, and inhibits glycation of collagen.

Another object of the present invention is to provide a compound represented by formula (I):

Another object of the present invention is to provide a compound represented by formula (I) or formula (II) for use in preparing an anti-glycation pharmaceutical composition.

In one embodiment of the present invention, the compounds of formula (I) and formula (II) are isolated from the aforementioned mango young fruit extract.

In one embodiment of the present invention, the compound of formula (I) or formula (II) inhibits collagen saccharification.

The invention discloses that the mango young fruit extract not only has ultraviolet protection and whitening effect on skin cells, but also can inhibit the saccharification of proteins and prevent the protein from losing its normal structure and function due to the saccharification reaction. In addition, the present invention also discloses a compound of formula (I) and formula (II) having significant anti-glycation activity. In view of ultraviolet radiation, excessive precipitation of melanin, and saccharification reactions, which can cause skin aging appearance, especially the saccharification reaction is one of the causes of body aging, and the mango fruit extracts and compounds disclosed in the present invention can be used to prepare skin anti-aging based on their characteristics. The composition of ultraviolet light and whitening and anti-glycation, therefore, the present invention provides a new countermeasure to delay the aging of the skin and even the individual. The aforementioned anti-ultraviolet and whitening and anti-glycation composition of the skin can be powder, granule, solution, colloid or paste, and can be made into medicine, food, drink, or nutritional supplement by oral administration, skin application or other methods Give a body.

The embodiments of the present invention will be further described below with reference to the drawings. The examples listed below are used to clarify the features and applications of the present invention, but not to limit the scope of the present invention. Within the spirit and scope of the present invention, some modifications and retouching can be done. Therefore, the protection scope of the present invention shall be determined by the scope of the attached patent application.

FIG. 1 shows the relative melanin content of melanoma cells B16F10 treated with kojic acid or young mango fruit extract according to an embodiment of the present invention.

FIG. 2 shows the cell viability of skin fibroblasts after pretreatment with young mango fruit extract according to an embodiment of the present invention and irradiation with ultraviolet A.

FIG. 3 shows the cell survival rate of skin fibroblasts after being irradiated with ultraviolet A and then treated with young mango fruit extract according to an embodiment of the present invention.

FIG. 4 shows the effect of young mango fruit extract on the production of collagen glycation end products according to an embodiment of the present invention.

Figure 5 shows the mass spectrum of Compound 1.

FIG. 6A shows a hydrogen nuclear magnetic resonance ( ¹ H-NMR) spectrum of Compound 1. FIG.

FIG. 6B shows a carbon nuclear magnetic resonance ( ¹³ C-NMR) spectrum of Compound 1. FIG.

FIG. 6C shows a hydrogen-hydrogen correlation spectroscopy (COSY) spectrum of Compound 1. FIG.

FIG. 6D shows a carbon-hydrogen heteronuclear single quantum correlation (HSQC) spectrum of Compound 1. FIG.

FIG. 6E shows a carbon-hydrogen heteronuclear multiple bond correlation (HMBC) spectrum of Compound 1. FIG.

FIG. 7 shows the effect of compounds of formula (I) and formula (II) on the production of collagen glycation end products.

definition

Unless otherwise stated, the terms "a," "an," and the like used herein are to be understood to include the singular and plural forms.

The values used in this article are approximate. All experimental data are shown in the range of 20%, preferably in the range of 10%, and most preferably in the range of 5%.

The pharmaceutical compositions described herein can be prepared into a dosage form suitable for parenterally or oral administration using techniques well known to those skilled in the art, including but not limited to : Injection [for example, sterile aqueous solution or dispersion], powder (sterile powder), tablet, troche, lozenge , Pill, capsule, dispersable powder, granule, solution, suspension, emulsion, syrup, elixir, concentrated Slurry and the like.

The pharmaceutical compositions described herein can be administered by parenteral routes, including but not limited to: intraperitoneal injection, subcutaneous injection, intramuscular injection, and Intravenous injection.

The pharmaceutical compositions described herein may include a pharmaceutically acceptable carrier that is widely used in pharmaceutical manufacturing technology. The pharmaceutically acceptable carrier may include one or more agents selected from the group consisting of a solvent, an emulsifier, a suspending agent, and a decomposer. , Binding agent, excipient, stabilizing agent, chelating agent, diluent, gelling agent, preservative, lubricant Lubricants, absorption delaying agents, liposomes, and the like. The selection and quantity of these reagents fall within the professional literacy and routine techniques of those skilled in the art.

The aforementioned pharmaceutically acceptable carrier includes a solvent selected from the group consisting of water, normal saline, phosphate buffered saline (PBS), a sugar-containing solution, Aqueous solution containing alcohol, and combinations thereof.

Materials and Methods

material

DMEM medium (Gibco Dulbecco's modified Eagle's medium), Eagle's minimum essential medium (Embedded MEM medium) containing Earle's balanced salt solution were purchased from Thermo Fisher Scientific, Gibco fetal bovine serum (FBS), penicillin / Streptomycin (Gibco penicillin / streptomycin), phosphate buffered saline (Gibco PBS), sodium pyruvate, sodium bicarbonate, and non-essential amino acids. 3- (4,5-dimethyl-2-thiazole) -2,5-diphenyltetrazolium bromide (3- (4,5-dimethylthiazol-2-yl) purchased from AMERSCO for cell survival analysis ) -2,5-diphenyltetrazolium bromide (MTT). Dimethyl sulfoxide (DMSO) was purchased from Echo chemical.

The solvent purchased from Taiwan Merck, including n-hexane (n -hexane), ethyl acetate (ethyl acetate), acetone (acetone), methanol (Methanol), ethanol (Ethanol), acetonitrile (acetonitrile), chloroform - d ₁ (Degree of deuteration 99.5%), methanol- d ₆ (degree of deuteration 99.5%), heavy water (deuterium oxide (degree of deuteration> 99.8%)), and dimethyl sulfide- d ₆ (dimethyl sulfoxide- d ₆ , deuterium Degree of transformation> 99.9%).

Chemical analysis instrument

The compound separation system uses column chromatography and thin layer chromatography (TLC). The medium pressure liquid chromatography (MPLC) system is Combi Flash ^® Rf + (Teledyne ISCO); the column system is selected from Sephadex LH-20 (Amersham Biosciences), Diaion HP-20 (Mitsubishi Chemical), Merck Kieselgel 60 (40-63 μm, Art.9385), and Merck LiChroprep® RP-18 (40-63 μm, Art.0250). High performance liquid chromatography (HPLC) system equipped with Hitachi L-2310 series pumps, Hitachi L-2420 UV-VIS detector (detection wavelength 200nm to 380nm), and D-2000 Elite data processing Software; The column system uses self-analytical Discovery® HS C ₁₈ (250 × 4.6mm, 5μm; SUpelCO) and Mightysil RP-18 GP 250 (250 × 4.6mm, 5μm; Kanto Chemical), and semi-preparation Discovery® HS C ₁₈ (250 × 10.0mm, 5μm; SUpelCO) and production-grade Discovery® HS C ₁₈ (250 × 21.0mm, 5μm; SUpelCO). The chromatography system is equipped with UV lamp UVP UVGL-25 (wavelengths 254nm and 365nm). Thin layer chromatography tablets are aluminum sheets coated with silicone 60 F ₂₅₄ (0.25mm; Merck) or RP-18 F _254S (0.25mm; Merck).

The chemical structure of the compound was determined by mass spectrometry (MS) and nuclear magnetic resonance spectrometry (NMR). Specifically, two-dimensional ion trap tandem Fourier transform mass spectrometry (Bruker amaZon SL system) and electrospray ionization tandem mass spectrometry (ESI-MS / MS; Thermo Scientific Orbitrap Elite system) were used; and 400 MHz Varian 400 FT-NMR was used to obtain -Dimensional and two-dimensional NMR spectra, using tetramethylsilane (TMS) as an internal standard (δ = 0).

Cell culture

The cells used in the following examples include mouse melanoma cell line B16F10 (ATCC CRL-6475) and human skin fibroblast CCD-966SK (ATCC CRL-1881) purchased from the American Type Culture Collection (ATCC). ). B16F10 cells were cultured under the conditions of 37 ° C and 5% carbon dioxide in a DMEM medium supplemented with 10% FBS and 1% penicillin / streptomycin, hereinafter referred to as DMEM cell culture medium. CCD-966SK cells were cultured at 37 ° C and 5% carbon dioxide with the addition of 10% FBS, 1 mM sodium pyruvate, 1.5 g / L sodium bicarbonate, 0.1 mM non-essential amino acids, and 1% penicillin / streptomycin MEM medium, hereinafter referred to as MEM cell culture medium.

Melanin production test

The method for measuring the melanin content of the melanoma cell line B16F10 is briefly described below. Cells were collected from the designated cell culture. The cells were washed with a PBS solution and treated with a trypsin solution for 3 minutes. The resulting suspended cells were collected by centrifugation (400 xg, 5 minutes), washed twice with the PBS solution, and then resuspended in 200 μL of the PBS solution. The cell suspension was frozen with liquid nitrogen for 10 minutes, and then left at room temperature for about 30 minutes to complete thawing, and the supernatant was removed by centrifugation (12,000 g, 3 minutes). The remaining cell pellet was mixed with 120 μL of sodium hydroxide and heated at 60 ° C. for 1 hour to obtain a cell lysate. 100 μL of the cell lysate was transferred to a 96-well plate, and the absorbance of the cell lysate at 450 nm (OD 450) was measured using an enzyme-linked immunosorbent assay reader (BioTek). The relative melanin content is calculated according to the following formula: relative melanin content (%) = (OD450 value of each group / OD450 value of blank control group) × 100%.

MTT analysis

(formazan)結晶。最終，使用ELISA讀盤機(BioTek)測量該細胞混合物在570nm的吸光值(OD 570)。細胞存活率係依下列公式計算：細胞存活率=(各組的OD 570值/空白對照組的OD 570值)×100%。 Cell viability was determined by MTT analysis. Briefly, MTT solution (4 mg / ml MTT in PBS solution) was added to cells in a 96-well plate at 15 μL / well, and reacted at room temperature for 4 hours. After removing the reaction solution, DMSO was added to the cells at 50 μL / well and the reaction was shaken for 10 minutes to dissolve the generated formazan

(formazan) Crystal. Finally, the absorbance (OD 570) of the cell mixture at 570 nm was measured using an ELISA disk reader (BioTek). Cell survival rate was calculated according to the following formula: Cell survival rate = (OD 570 value of each group / OD 570 value of blank control group) x 100%.

Statistical Analysis

Data are expressed as mean ± standard deviation (SD). Statistical analysis was performed using Excel software, and statistically significant differences between the data were determined by student's t-test.

Example 1

Preparation of Mango Young Fruit Extract

The mango indica described in this article refers to the mango variety produced in Taiwan, but not limited to this. Generally speaking, the growth and development of mango fruit is divided into four stages, which are (1) the young fruit stage: after flowering, the mango begins to grow slowly and is green; (2) the rapid growth period: the fruit grows rapidly, Starch accumulates gradually; (3) Ripening period: The fruit's inner peel hardens and then enters the ripening period. At this time, the appearance of the fruit changes little but the weight of the fruit continues to increase. Some physical and chemical changes are still ongoing, such as fruit Decreased firmness, increased sugar content, and yellowed peels make the fruits close to the ripe and edible stage; (4) Aging period: the fruits begin to age as soon as they are ripe. The young mango or immature mango fruit described herein refers to a mango fruit that has not entered the mature stage and has not turned yellow.

To obtain a young mango fruit extract, an unripe mango fruit having a length of about 3 to 7 cm is ground with a homogenizer. Thereafter, water, alcohols, or a mixture of alcohol and water is used as a solvent to extract the homogenate of young mango fruit, and the solvent can add a mixed acid of 0.1% to 5% organic acids (such as acetic acid and citric acid) and hydrochloric acid. Wherein the weight ratio of the solvent to the homogenate of the young mango fruit is 20: 1 to 1: 1. The extraction temperature is between 55 ° C and 100 ° C, preferably between 55 ° C and 85 ° C. The mango fruit extracts described in Examples 2-5 below are all extracted with an aqueous solution containing 0.1 to 0.5% acetic acid, citric acid and hydrochloric acid, and the extraction time is 0.5 to 3 hours.

After the mango fruit young fruit extract obtained through the above extraction step is cooled to room temperature, a supernatant can be further centrifuged at a speed of 3000 to 5000 rpm at room temperature for 5 to 10 minutes, and the supernatant can be used with 400 mesh ( mesh) to remove residual solids. The filtered young mango fruit extract can be further concentrated under reduced pressure at 45 ° C to 70 ° C to obtain a concentrated product. In order to obtain a solid young mango fruit extract, the aforementioned concentrated young mango fruit extract may be dried in a dry manner such as freeze-drying, spray-drying or the like to remove the solvent, thereby obtaining a powdery young mango fruit extract.

Example 2

Mango fruit extract inhibits melanin formation

In order to test the effect of young mango fruit extract on melanin production, the melanoma cell line B16F10 was measured by the melanoma cell line B16F10 treated with the young mango fruit extract described in Example 1 to change the melanin content. Briefly, B16F10 cells were seeded at 1.5 × 10 ⁵ cells / well into a 6-well culture plate, each well containing 3 mL of DMEM cell culture medium. After culturing the cells at 37 ° C for 24 hours, the cell culture medium was removed, and 3 mL of DMEM cell culture medium containing 0.25 mg / mL young mango fruit extract (experimental group) or 0.25 mg / mL gallic acid (positive control group) was added to Cells in each well. A blank control group was set up to treat cells with only 3 mL of DMEM cell culture medium. After 48 hours of incubation at 37 ° C, the aforementioned three groups of cells were collected to determine the melanin content (triplicate test).

Figure 1 shows the relative melanin content of the three groups of melanoma cells. According to the figure, compared with the blank control group, the administration of young mango fruit extract significantly reduced the relative black content by about 18%, and the effect of suppressing the formation of this black pigment was equivalent to that of the same concentration of osmic acid (a conventional whitening compound). This result indicates that the mango fruit extract has no less melanin formation inhibitory effect than the conventional whitening compounds.

Example 3

Mango fruit extract prevents and reduces skin fibroblast death caused by ultraviolet A radiation

In order to test the effect of young mango fruit extracts on protecting the skin against ultraviolet radiation, human skin fibroblasts were evaluated by cell survival analysis (MTT analysis). CCD-966SK was previously treated with the young mango fruit extracts described in Example 1 and then irradiated with ultraviolet A ( UVA protection test), or the survival rate of cells treated with the young mango fruit extract (UVA damage repair test) before being irradiated with ultraviolet A.

3.1 UVA protection test

CCD-966SK cells were seeded into 96-well culture plates at 5 × 10 ³ cells / well, and each well contained 200 μL of MEM cell culture medium. After culturing the cells at 37 ° C for 24 hours, remove the cell culture medium, add 200 μL of MEM cell culture medium without or containing 0.5 mg / mL young mango fruit extract to each well cell, and then culture the cells at 37 ° C for 24 hours. The aforementioned cells were treated in the following manner: (a) cells not pretreated with young mango fruit extract were cultured for 1 hour in the absence of ultraviolet radiation (blank control group); (b) cells not pretreated with young mango fruit extract In a UV irradiation box (Vilber), irradiate with 15J / cm ² UVA for 1 hour (UVA group), the radiation dose will cause half of the cell death; or (c) cells pretreated with young mango fruit extract at 15J / cm ² UVA irradiation for 1 hour (mango young fruit extract + UVA group). Thereafter, MTT analysis was performed on each group of cells to calculate the cell survival rate.

Figure 2 shows the cell viability of skin fibroblasts after different treatments. According to Figure 2, compared with the blank control group, the cell survival rate of the UVA group was about 61.8%, showing that ultraviolet A radiation can cause a large number of skin fibroblasts to die. Compared with the UVA group, the pretreatment of young mango fruit extracts increased the cell survival rate to about 84.1% under UV irradiation. This result indicates that the application of young mango fruit extract can prevent the damage or death of skin cells caused by ultraviolet rays.

3.2 UVA damage repair test

CCD-966SK cells were seeded into 96-well culture plates at 5 × 10 ³ cells / well, and each well contained 200 μL of MEM cell culture medium. After culturing the cells at 37 ° C for 24 hours, treat the cells in each well and replace the cell culture medium in the following manner: (a) After incubation for 1 hour in the absence of ultraviolet radiation, 200 μL of MEM cell culture medium containing no mango young fruit extract was applied (blank control) Group); (b) irradiated with 15 J / cm ² ultraviolet A for 1 hour, and applied 200 μL of MEM cell culture medium (UVA group) without young mango fruit extract; or (c) irradiated with 15 J / cm ² ultraviolet A for 1 hour Then, 200 μL of MEM cell culture medium (UVA + mango fruit extract group) containing 0.5mg / mL young mango fruit extract was applied. Cells of each group were cultured at 37 ° C for 24 hours, and MTT analysis was performed to calculate the cell survival rate.

Figure 3 shows the cell viability of skin fibroblasts after different treatments. According to Fig. 3, compared with the blank control group, the cell survival rate of the UVA group was about 42.9%, but the cells were treated with young mango fruit extract after UV irradiation to increase the survival rate to about 78.4%. This result indicates that the application of young mango fruit extract can reduce or repair the damage of skin cells caused by ultraviolet rays.

Example 4

Mango fruit extract inhibits collagen saccharification

In order to test the anti-glycation activity of the young mango fruit extract, the anti-glycation analysis was used to determine the inhibitory effect of the young mango fruit extract described in Example 1 at different concentrations on the glycation reaction of pig collagen. In brief, a 200 mM phosphate buffer solution (pH 7.4) was first used to prepare a 60 mg / mL collagen solution (containing 0.06% sodium azide) and a 1.5 M fructose solution. For collagen saccharification, a mixture of 0.2 mL of collagen solution and 0.2 mL of fructose solution and 0.2 mL of 140, 70, 14, 1.4, or 0.14 mg / mL young mango fruit extract samples (diluted with deionized water) ) Or deionized water (blank control group), mix uniformly, and react at 50 ° C. for 24 hours, and then add aminoguanidine (AG, purchased from Sigma) to stop the saccharification reaction. A spectrofluorimeter (FLx 800, BioTek) was used to measure the fluorescence intensity (excitation wavelength 360nm, detection wavelength 460nm) of the reaction solution (0.1mL) at 0 hours and 24 hours, and the collagen glycosylation end product was calculated according to the following formula (AGEs) generation rate: AGEs generation rate = [(sample fluorescence intensity _{24 hours} -sample fluorescence intensity _{0 hours} ) / (blank control group fluorescence intensity _{24 hours} -blank control group fluorescence intensity _{0 hours} )] x 100% .

Figure 4 shows the effect of young mango fruit extract on the production of collagen glycosylation end products. According to the figure, compared with the blank control group, the treatment of 140, 70, 14, 1.4, and 0.14 mg / mL young mango fruit extracts significantly reduced the final saccharification product yield to about 3.6%, 9.6%, 39.7%, and 59.2, respectively. % And 62.9%, showing that young mango fruit extract can inhibit the glycation reaction of protein in the body, and then delay the aging of the individual.

Example 5

Preparation and identification of compounds of formula (I) and formula (II)

In order to obtain the anti-glycation active ingredient in young mango fruit extract, first, 1 L of young mango fruit extract was prepared according to the method described in Example 1, and the extract was extracted three times by means of liquid phase distribution of ethyl acetate and water in equal proportions. . The obtained ethyl acetate layer extracts were combined and concentrated under reduced pressure to obtain about 4.7 g of an ethyl acetate layer extract.

According to a bioassay guided fractionation method, column chromatography was performed on the ethyl acetate layer extract (approximately 4.7 g) using a Diaion HP-20 column and an eluent with a decreasing gradient of water and methanol. , To obtain 3 divided layers (labeled as F1 to F3 respectively). Thereafter, column chromatography was performed on the F2 or F3 layer using Sephadex LH-20 column and methanol eluent, and the resulting effluent was separated by thin-layer chromatography to obtain three separate layers (labeled separately) (F2-1 to F2-3 and F3-1 to F3-3). The F3-2 layer was further subjected to high performance liquid chromatography using a mixed solution of water and methanol at a volume ratio of 1: 1 as a mobile phase, and about 13.0 mg of Compound 1 was isolated. In addition, the F2-1 divided layer was further subjected to high-performance liquid chromatography using a mixture of water and methanol at a volume ratio of 2: 1 as a mobile phase, and about 23.0 mg of Compound 2 was isolated.

The chemical structures of Compound 1 and Compound 2 were determined by mass spectrometry and nuclear magnetic resonance spectroscopy (NMR). The compound is a pale brown oil, viewed from the spectrum shown in FIG. 5 to a pseudo-molecular ion peak of ^{m / z 907 [MH] -} , it is speculated that the molecular weight of Compound 1 was 908Da. According to the hydrogen nuclear magnetic resonance spectrum shown in FIG. 6A, Compound 1 has a main structure composed of one sugar group and five aromatic rings. According to the carbon nuclear magnetic resonance spectrum shown in FIG. 6B, Compound 1 has a total of 41 carbon absorption signals, which are 5 carbonyl absorption signals, 5 benzene ring absorption signals, and 1 group of sugar-based absorption signals. In addition, according to the two-dimensional nuclear magnetic resonance spectra (COSY, HSQC, HMBC) shown in Figs. 2C to 2E, it is possible to infer the connection manner and the position of the substituents of the single sugar group and the five aromatic rings in Compound 1, so it is confirmed that Compound 1 is a kind of Hydrolyzed tannin, which has a structure represented by formula (I). Compound 2 was confirmed by literature comparison with its mass spectrum and nuclear magnetic resonance spectrum to be a hydrolyzed tannin, which has a structure as shown in formula (II).

Example 6

Anti-glycation activity of compounds of formula (I) and formula (II)

In order to test the anti-glycation activity of the compounds of formula (I) and formula (II), the anti-glycation analysis as described in Example 4 was used to determine the inhibitory effect of these compounds on the glycation reaction of porcine collagen. In short, a mixture of 0.2 mL of a collagen solution and 0.2 mL of a fructose solution and 0.2 mL of a 100 μg / mL sample of an aqueous solution of a compound of formula (I) or formula (II) or deionized water (blank control group) are uniformly mixed at 50 The reaction was performed at a temperature of 24 ° C for 24 hours, and the fluorescence intensity of the reaction solution (0.1 mL) at 0 hours and 24 hours was measured to calculate the collagen glycosylation end product production rate.

FIG. 7 shows the effect of compounds of formula (I) and formula (II) on the production of collagen glycation end products. According to the figure, compared with the blank control group, the treatment of the compounds of formula (I) and formula (II) significantly reduced the amount of end-saccharification products to about 60.2% and 73.7%, respectively, showing that the two compounds have significant anti-glycation activity.

In summary, the present invention discloses that the mango young fruit extract not only has ultraviolet protection and whitening effect on skin cells, but also can inhibit the saccharification of proteins and prevent the protein from losing its normal structure and function due to the saccharification reaction. In addition, the present invention also discloses a compound of formula (I) and formula (II) having significant anti-glycation activity. In view of ultraviolet radiation, excessive precipitation of melanin, and saccharification reactions, which can cause skin aging appearance, especially the saccharification reaction is one of the causes of body aging, and the mango fruit extracts and compounds disclosed in the present invention can be used to prepare skin anti-aging based on their characteristics. The composition of ultraviolet light and whitening and anti-glycation, therefore, the present invention provides a new countermeasure to delay the aging of the skin and even the individual. The aforementioned anti-ultraviolet, whitening and anti-glycation composition can be powder, granule, solution, colloid or paste, and can be made into medicine, food, drink, or nutritional supplement, and can be administered by oral, skin application or other methods A body.

Claims (10)

    A young mango fruit extract is used for preparing skin anti-ultraviolet and whitening and anti-glycation pharmaceutical composition. The young mango fruit extract is obtained by extracting a young mango fruit with a solvent, wherein the length of the young mango fruit is 3 to 7 cm of unripe mango fruit.         The use as described in item 1 of the patent application range, wherein the weight ratio of the solvent to the young mango fruit ranges from 20: 1 to 1: 1.         The use as described in item 1 of the patent application range, wherein the extraction is performed at 55 ° C to 100 ° C.         The use as described in item 1 of the scope of patent application, wherein the solvent is water.         The use as described in item 1 of the patent application range, wherein the concentration of the young mango fruit extract is at least 0.25 mg / mL.         The use as described in item 1 of the scope of patent application, wherein the young mango fruit extract prevents and repairs the damage of skin fibroblasts caused by ultraviolet A.         The use according to item 1 of the patent application scope, wherein the mango young fruit extract inhibits collagen saccharification.     A compound represented by formula (I):

Use of a compound shown below for the preparation of an anti-glycation pharmaceutical composition:

or

    The use according to item 9 of the scope of patent application, wherein the compound of formula (I) or formula (II) inhibits collagen saccharification.