An active compound of Lycopodiastrum cernuum, its preparation method and application

Abstract

The present invention belongs to the technical field of plant extracts, and specifically discloses an active compound of Lycopodiastrum cernuum, a preparation method thereof, and an application thereof. The structure of the compound includes: wherein, I is phyllocladanol; II is lycocladinol; III is 2,4-di-tert-butylphenol; IV is 5-hydroxymethylfurfural; V is 5-ethoxymethylfurfural; VI is ethyl palmitate; VII is ethyl linoleate; VIII is 1-eicosene. By adopting the active compound of Lycopodiastrum cernuum, the preparation method thereof, and the application thereof, the present invention discovers a new compound, the tyrosinase activity of which is equivalent to that of the positive control, and solves the technical problems that existing whitening substances have risks such as allergy and carcinogenesis and are prone to discoloration under the influence of the environment, thereby affecting the whitening effect; the active compound of Lycopodiastrum cernuum with good whitening effect is obtained by a simple extraction method.

Description

Technical Field

[0001] The present invention belongs to the technical field of plant extracts, and particularly relates to an active compound of Palhinhaea cernua, a preparation method thereof, and an application thereof. Background Art

[0002] Tyrosinase plays many different roles in humans, insects, plants, and microorganisms. In humans, tyrosinase is involved in the formation of melanin. The melanin catalyzed and generated by tyrosinase determines the colors of the skin, hair, and iris in humans, and can also protect the skin from harmful effects such as ultraviolet (UV) radiation, oxidative stress, and DNA damage. However, the excessive accumulation of melanin caused by the abnormal expression of tyrosinase will cause pigmentation diseases, such as melanoma, melasma, and freckles. With the continuous deepening of the research on tyrosinase, more and more tyrosinase inhibitors have been developed and studied. Natural compounds are an important source of tyrosinase inhibitors. In the current cosmetics market, most whitening products take tyrosinase activity inhibitors as the main ingredient. Most whitening substances, such as hydroquinone and vitamin C, achieve whitening effects by inhibiting tyrosinase or antioxidant activity. Although hydroquinone has obvious whitening effects, it is prohibited from use due to carcinogenic risks. And vitamin C is prone to discoloration due to factors such as light, heat, humidity, and pH value during the preparation process. There is no report on the use of Palhinhaea cernua in whitening products in the current market.

[0003] Palhinhaea cernua is a plant of the genus Palhinhaea in the family Lycopodiaceae. It is a medium-sized to large-sized terrestrial plant, which is produced in the understory, forest edge, and shaded areas under shrubs or on rocks at an altitude of 100 - 1800 meters in Zhejiang, Jiangxi, Fujian, Taiwan, Hunan, Guangdong, Hong Kong, Guangxi, Hainan, Sichuan, Chongqing, Guizhou, Yunnan, etc. According to "Chinese Herbal Medicine", the whole herb of Palhinhaea cernua is used as medicine. It is bitter in taste and neutral in nature, and enters the liver, spleen, and kidney meridians, and has the effects of dispelling wind and dampness, relaxing tendons and activating blood circulation, relieving cough and diuretic. Modern research shows that Palhinhaea cernua mainly contains chemical components such as alkaloids, triterpenoids, and flavonoids, and has effects such as anti-inflammatory, cytotoxic, acetylcholinesterase, α-glucosidase, and xanthine oxidase. Therefore, Palhinhaea cernua has broad application prospects in complementary and alternative medicines. However, up to now, no research report on the tyrosinase activity of Palhinhaea cernua has been found.

[0004] Therefore, there is a need in the art to develop an active compound of Palhinhaea cernua, a preparation method thereof, and an application thereof, which can effectively solve the technical problems that existing whitening substances have risks such as allergy and carcinogenesis, and are prone to discoloration under the influence of the environment, thus affecting the whitening effect. Summary of the Invention

[0005] The object of the present invention is to provide an active compound of Lycopodium cernuum and its preparation method and application. A new compound has been discovered, and its tyrosinase activity is equivalent to that of the positive control, solving the technical problems that existing whitening substances have risks such as allergies and carcinogens, and are prone to discoloration under the influence of the environment, thus affecting the whitening effect; an active compound of Lycopodium cernuum with good whitening effect is obtained by a simple extraction method.

[0006] To achieve the above object, the present invention provides an active compound of Lycopodium cernuum, and the structure of the compound includes:

[0007]

[0008] Among them, I is phyllocladanol; II is lycocladoniol; III is 2,4-di-tert-butylphenol; IV is 5-hydroxymethylfurfural; V is 5-ethoxymethylfurfural; VI is ethyl palmitate; VII is ethyl linoleate; VIII is 1-eicosen.

[0009] A preparation method of an active compound of Lycopodium cernuum includes the following steps:

[0010] Step S1: Naturally air-dry and crush the Lycopodium cernuum sample to obtain a powder;

[0011] Step S2: Soak the powder obtained in Step S1 with absolute ethanol, and concentrate under reduced pressure to obtain an ethanol extract;

[0012] Step S3: After dispersing the ethanol extract obtained in Step S2, extract with petroleum ether and chloroform in sequence to obtain a petroleum ether extract and a chloroform extract;

[0013] Step S4: Separate the petroleum ether extract or chloroform extract obtained in Step S3 on a silica gel chromatography column with 300-400 mesh, and elute with a petroleum ether-ethyl acetate mobile phase to obtain multiple components; then further separate and purify, and through an elution program, an active compound of Lycopodium cernuum is obtained.

[0014] Preferably, in Step S2, the powder obtained in Step S1 is soaked with absolute ethanol 3 times, and each soaking lasts for 7 days; the volume ratio of absolute ethanol to the powder is 1:1.

[0015] Preferably, in Step S2, the temperature for concentration under reduced pressure is 37°C, the vacuum degree is 0.1 MPa, the rotation speed is 70 rpm / min, and the temperature of the condensed water is 5°C.

[0016] Preferably, in Step S4, the silica gel chromatography column is one or both of a 300-400 mesh silica gel chromatography column or a Sephadex LH-20 gel column chromatography.

[0017] Preferably, in step S4, in the elution procedure, the mobile phase is one or more of petroleum ether - ethyl acetate, dichloromethane - methanol, and petroleum ether - dichloromethane - methanol.

[0018] Use of an active compound of Lycopodiastrum cernuum in the preparation of a tyrosinase inhibitor.

[0019] Use of an active compound of Lycopodiastrum cernuum in the preparation of skin care products.

[0020] The present invention adopts the above - mentioned active compound of Lycopodiastrum cernuum, its preparation method and application, and the beneficial effects are as follows:

[0021] (1) Applying the active compound in the Lycopodiastrum cernuum of the present invention to the preparation of skin - whitening skin care products has a relatively good inhibitory effect on tyrosinase activity.

[0022] (2) The present invention discovers a new compound, and its tyrosinase activity is equivalent to that of the positive control, solving the technical problems of existing skin - whitening substances such as allergy, carcinogenicity and other risks, and being prone to color change under the influence of the environment, thus affecting the skin - whitening effect.

[0023] (3) The present invention provides a simple extraction method. Dry, crush the whole herb of Lycopodiastrum cernuum, soak it in absolute ethanol, and concentrate it under reduced pressure to obtain an extract. After dispersing the ethanol extract, extract it successively with petroleum ether and chloroform to obtain a petroleum ether extract and a chloroform extract, and elute the extracts to further extract the active compound of Lycopodiastrum cernuum with good skin - whitening effect.

[0024] The following further describes the technical solution of the present invention in detail through the drawings and examples. Description of the Drawings

[0025] Figure 1 It is the preparation flow chart of an embodiment of an active compound of Lycopodiastrum cernuum, its preparation method and application of the present invention. Detailed Embodiments

[0026] The following further illustrates the technical solution of the present invention through the drawings and examples.

[0027] Unless otherwise defined, the technical terms or scientific terms used in the present invention should have the ordinary meaning understood by those of ordinary skill in the field to which the present invention belongs.

[0028] Example 1

[0029] As Figure 1 shown, a preparation method of an active compound of Lycopodiastrum cernuum includes the following steps:

[0030] Step S1: Naturally air - dry and crush 450 g of Lycopodiastrum cernuum samples to obtain powder.

[0031] Step S2: Soak the powder obtained in Step S1 with absolute ethanol, with the volume ratio of absolute ethanol to the powder being 1:1. Soak it 3 times, with each soaking for 7 days, at a temperature of 37°C, a vacuum degree of 0.1 MPa, a rotation speed of 70 rpm / min, and concentrate it under reduced pressure at a condensing water temperature of 5°C to obtain 415 g of ethanol extract.

[0032] Step S3: After dispersing the ethanol extract obtained in Step S2, extract it successively with petroleum ether and chloroform to obtain petroleum ether extract and chloroform extract.

[0033] Step S4: Separate the chloroform extract obtained in Step S3 on a silica gel column chromatography column with a mesh size of 300 - 400, and elute it with a mobile phase of petroleum ether - ethyl acetate according to the gradients of 20:1, 10:1, 8:2, 7:3, 1:1, and 0:1 to obtain 6 components (Fr.C1 - Fr.C6).

[0034] Separate the component Fr.C5 on a silica gel column chromatography column with a mesh size of 300 - 400, and elute it with a mobile phase of petroleum ether - ethyl acetate according to the gradients of 8:2, 7:3, 6:4, and 0:1 to obtain 20 sub - components (Fr.C5A - Fr.C5T).

[0035] Separate the sub - component Fr.C5S on a silica gel column chromatography column with a mesh size of 300 - 400, elute it with a mobile phase of petroleum ether - ethyl acetate according to 1:1, and then perform column chromatography on a SephadexLH - 20 gel column, and elute it with a mobile phase of petroleum ether - dichloromethane - methanol according to 2:1:1, and repeat the elution in multiple rounds to obtain the active compound I of Palhinhaea cernua, and its structural formula is:

[0036]

[0037] Example 2

[0038] A preparation method of the active compound of Palhinhaea cernua, comprising the following steps:

[0039] Step S1: Naturally air - dry and pulverize 450 g of Palhinhaea cernua samples to obtain powder.

[0040] Step S2: Soak the powder obtained in Step S1 with absolute ethanol, with the volume ratio of absolute ethanol to the powder being 1:1. Soak it 3 times, with each soaking for 7 days, at a temperature of 37°C, a vacuum degree of 0.1 MPa, a rotation speed of 70 rpm / min, and concentrate it under reduced pressure at a condensing water temperature of 5°C to obtain 415 g of ethanol extract.

[0041] Step S3: After dispersing the ethanol extract obtained in Step S2, extract it successively with petroleum ether and chloroform to obtain petroleum ether extract and chloroform extract.

[0042] Step S4: The chloroform extract obtained in Step S3 was separated on a silica gel column chromatography with a mesh size of 300 - 400, and eluted with a mobile phase of petroleum ether - ethyl acetate according to the gradients of 20:1, 10:1, 8:2, 7:3, 1:1, and 0:1 to obtain 6 components (Fr.C1 - Fr.C6).

[0043] Component Fr.C5 was separated on a silica gel column chromatography with a mesh size of 300 - 400, and eluted with a mobile phase of petroleum ether - ethyl acetate according to the gradients of 8:2, 7:3, 6:4, and 0:1 to obtain 20 sub - components (Fr.C5A - Fr.C5T).

[0044] Sub - component Fr.C5L was separated on a silica gel column chromatography with a mesh size of 300 - 400, and eluted with a mobile phase of dichloromethane - methanol according to the ratio of 100:1 to obtain the active compound II of Palhinhaea cernua, and its structural formula is:

[0045]

[0046] Example 3

[0047] A preparation method of an active compound of Palhinhaea cernua, comprising the following steps:

[0048] Step S1: 450 g of Palhinhaea cernua samples were naturally air - dried and pulverized to obtain powder.

[0049] Step S2: The powder obtained in Step S1 was soaked with absolute ethanol, and the volume ratio of absolute ethanol to powder was 1:1. Soak it 3 times, soak for 7 days each time, at a temperature of 37 °C, a vacuum degree of 0.1 MPa, a rotation speed of 70 rpm / min, and concentrate under reduced pressure at a condensed water temperature of 5 °C to obtain 415 g of ethanol extract.

[0050] Step S3: After the ethanol extract obtained in Step S2 was dispersed, it was successively extracted with petroleum ether and chloroform to obtain petroleum ether extract and chloroform extract.

[0051] Step S4: The chloroform extract obtained in Step S3 was separated on a silica gel column chromatography with a mesh size of 300 - 400, and eluted with a mobile phase of petroleum ether - ethyl acetate according to the gradients of 20:1, 10:1, 8:2, 7:3, 1:1, and 0:1 to obtain 6 components (Fr.C1 - Fr.C6).

[0052] Component Fr.C4 was separated on a silica gel column chromatography with a mesh size of 300 - 400, and eluted with a mobile phase of petroleum ether - ethyl acetate according to the ratio of 6:1 to obtain 3 sub - components (Fr.C4A - Fr.C4C).

[0053] The sub-component Fr.C4A was subjected to Sephadex LH-20 gel column chromatography, eluted with a mobile phase of petroleum ether-dichloromethane-methanol in a ratio of 2:1:1, and then subjected to silica gel column chromatography with 300-400 mesh, eluted with a mobile phase of petroleum ether-dichloromethane in a ratio of 5:1 to obtain the active compound III of Lycopodium cernuum L., and its structural formula is:

[0054]

[0055] Example 4

[0056] A preparation method of an active compound of Lycopodium cernuum L. comprises the following steps:

[0057] Step S1: Naturally air-dry and crush 450 g of Lycopodium cernuum L. sample to obtain a powder.

[0058] Step S2: Soak the powder obtained in Step S1 with absolute ethanol, and the volume ratio of absolute ethanol to the powder is 1:1. Soak it 3 times, each time for 7 days, at a temperature of 37 °C, a vacuum degree of 0.1 MPa, a rotation speed of 70 rpm / min, and concentrate under reduced pressure at a condensing water temperature of 5 °C to obtain 415 g of ethanol extract.

[0059] Step S3: After dispersing the ethanol extract obtained in Step S2, extract it successively with petroleum ether and chloroform to obtain a petroleum ether extract and a chloroform extract.

[0060] Step S4: Separate the chloroform extract obtained in Step S3 on a silica gel chromatography column with 300-400 mesh, and elute it with a mobile phase of petroleum ether-ethyl acetate in gradients of 20:1, 10:1, 8:2, 7:3, 1:1, and 0:1 to obtain 6 components (Fr.C1-Fr.C6).

[0061] Component Fr.C6 was separated on a silica gel column chromatography column with 300-400 mesh, and eluted with a mobile phase of dichloromethane-methanol in ratios of 200:1, 40:1, 10:1, and 1:1 to obtain 7 sub-components (Fr.C6A-Fr.C6G).

[0062] The sub-component Fr.C6G was subjected to Sephadex LH-20 gel column chromatography, eluted with a mobile phase of petroleum ether-dichloromethane-methanol in a ratio of 2:1:1, and then subjected to silica gel column chromatography with 300-400 mesh, eluted with a mobile phase of dichloromethane-methanol in a ratio of 40:1 to obtain the active compound IV of Lycopodium cernuum L., and its structural formula is:

[0063]

[0064] Example 5

[0065] A preparation method of an active compound of Lycopodium cernuum L. comprises the following steps:

[0066] Step S1: Naturally air-dry and pulverize 450 g of the Palhinhaea cernua sample to obtain a powder.

[0067] Step S2: Soak the powder obtained in Step S1 with absolute ethanol at a volume ratio of absolute ethanol to powder of 1:1. Soak it 3 times, with each soaking for 7 days, at a temperature of 37°C, a vacuum degree of 0.1 MPa, a rotation speed of 70 rpm / min, and perform decompression concentration at a condensed water temperature of 5°C to obtain 415 g of an ethanol extract.

[0068] Step S3: After dispersing the ethanol extract obtained in Step S2, sequentially extract with petroleum ether and chloroform to obtain a petroleum ether extract and a chloroform extract.

[0069] Step S4: Subject the petroleum ether extract obtained in Step S3 to silica gel column chromatography with a mesh size of 300 - 400, and elute with a mobile phase of petroleum ether - ethyl acetate according to gradients of 0:1, 20:1, 10:1, 8:2, 1:1 to obtain 7 components (Fr.P1 - Fr.P7).

[0070] Subject the sub-component Fr.P4 to silica gel column chromatography with a mesh size of 300 - 400, and elute with a mobile phase of dichloromethane - methanol according to a ratio of 100:1 to obtain the Palhinhaea cernua active compound V, whose structural formula is:

[0071]

[0072] Example Six

[0073] A preparation method of a Palhinhaea cernua active compound, comprising the following steps:

[0074] Step S1: Naturally air-dry and pulverize 450 g of the Palhinhaea cernua sample to obtain a powder.

[0075] Step S2: Soak the powder obtained in Step S1 with absolute ethanol at a volume ratio of absolute ethanol to powder of 1:1. Soak it 3 times, with each soaking for 7 days, at a temperature of 37°C, a vacuum degree of 0.1 MPa, a rotation speed of 70 rpm / min, and perform decompression concentration at a condensed water temperature of 5°C to obtain 415 g of an ethanol extract.

[0076] Step S3: After dispersing the ethanol extract obtained in Step S2, sequentially extract with petroleum ether and chloroform to obtain a petroleum ether extract and a chloroform extract.

[0077] Step S4: Subject the chloroform extract obtained in Step S3 to silica gel column chromatography with a mesh size of 300 - 400, and elute with a mobile phase of petroleum ether - ethyl acetate according to gradients of 20:1, 10:1, 8:2, 7:3, 1:1, 0:1 to obtain 6 components (Fr.C1 - Fr.C6).

[0078] The active compound of Palhinhaea cernua was separated by silica gel column chromatography with 300 - 400 mesh on component Fr.C2. Petroleum ether - ethyl acetate mobile phase was used for elution in sequence according to 40:1 and 20:1. Sephadex LH - 20 gel column chromatography was carried out, and petroleum ether - dichloromethane - methanol mobile phase was used for elution according to 2:1:1 to obtain the active compound VI of Palhinhaea cernua, and its structural formula is:

[0079]

[0080] Example VII

[0081] A preparation method of the active compound of Palhinhaea cernua includes the following steps:

[0082] Step S1: Naturally air - dry and crush 450 g of Palhinhaea cernua sample to obtain powder.

[0083] Step S2: Soak the powder obtained in step S1 with absolute ethanol, and the volume ratio of absolute ethanol to powder is 1:1. Soak it 3 times, each time for 7 days, at a temperature of 37 °C, a vacuum degree of 0.1 MPa, a rotation speed of 70 rpm / min, and carry out reduced - pressure concentration at a condensing water temperature of 5 °C to obtain 415 g of ethanol extract.

[0084] Step S3: After dispersing the ethanol extract obtained in step S2, extract it with petroleum ether and chloroform in sequence to obtain petroleum ether extract and chloroform extract.

[0085] Step S4: Subject the chloroform extract obtained in step S3 to silica gel column chromatography with 300 - 400 mesh, and use petroleum ether - ethyl acetate mobile phase for gradient elution according to 20:1, 10:1, 8:2, 7:3, 1:1, 0:1 to obtain 6 components (Fr.C1 - Fr.C6).

[0086] The component Fr.C2 was separated by silica gel column chromatography with 300 - 400 mesh. Petroleum ether - ethyl acetate mobile phase was used for elution in sequence according to 40:1 and 20:1. Sephadex LH - 20 gel column chromatography was carried out, and petroleum ether - dichloromethane - methanol mobile phase was used for elution according to 2:1:1 to obtain the active compound VII of Palhinhaea cernua, and its structural formula is:

[0087]

[0088] Example VIII

[0089] A preparation method of the active compound of Palhinhaea cernua includes the following steps:

[0090] Step S1: Naturally air - dry and crush 450 g of Palhinhaea cernua sample to obtain powder.

[0091] Step S2: Soak the powder obtained in Step S1 with absolute ethanol at a volume ratio of absolute ethanol to powder of 1:1. Soak it three times, with each soaking lasting for 7 days, at a temperature of 37°C, a vacuum degree of 0.1 MPa, a rotation speed of 70 rpm / min, and concentrate it under reduced pressure at a condensed water temperature of 5°C to obtain 415 g of ethanol extract.

[0092] Step S3: After dispersing the ethanol extract obtained in Step S2, extract it successively with petroleum ether and chloroform to obtain a petroleum ether extract and a chloroform extract.

[0093] Step S4: Subject the chloroform extract obtained in Step S3 to silica gel column chromatography with a mesh size of 300 - 400, and elute it with a mobile phase of petroleum ether - ethyl acetate at gradients of 20:1, 10:1, 8:2, 7:3, 1:1, and 0:1 to obtain 6 components (Fr.C1 - Fr.C6).

[0094] Subject the component Fr.C4 to silica gel column chromatography with a mesh size of 300 - 400 and elute it with a mobile phase of petroleum ether - ethyl acetate at a ratio of 6:1 to obtain 3 sub - components Fr.C4A - Fr.C4C.

[0095] Subject the sub - component Fr.C4A to silica gel column chromatography with a mesh size of 300 - 400, and elute it with a mobile phase of petroleum ether - ethyl acetate at ratios of 50:1, 20:1, and 7:1 to obtain the active compound VIII of Palhinhaea cernua, and its structural formula is:

[0096]

[0097] Verify the whitening effects of the active compounds I - VIII of Palhinhaea cernua prepared in the examples through experiments:

[0098] (1) Preparation of L - dopa solution: Take 100.0 mg of dry L - dopa powder, dissolve it with phosphate buffer solution with a pH of 6.8, transfer it to a 100 mL brown volumetric flask, and make up the volume to the scale to obtain a 1.0 mg / mL L - dopa solution.

[0099] (2) Preparation of tyrosinase solution: Take 2 mg of tyrosinase powder, dissolve it with phosphate buffer solution with a pH of 6.8, transfer it to a 100 mL brown volumetric flask, and make up the volume to the scale to obtain 100 U / mL.

[0100] (3) Preparation of reference substance solution: Weigh 1.0 mg of vitamin C powder, add an appropriate amount of absolute ethanol to dissolve it, transfer it to a 10 mL brown volumetric flask, and make up the volume to the scale line to obtain a 1.0 mg / mL vitamin C stock solution, and store it at room temperature for later use.

[0101] (4) Preparation of test sample solution: Prepare it into a 1.0 mg / mL solution for later use.

[0102] (5) The solution of the sample to be tested and the experimental conditions for tyrosinase activity inhibition are shown in Table 1.

[0103] Table 1 Reaction system for tyrosinase activity inhibition experiment

[0104] Reaction system PBS solution Sample solution Tyrosinase solution L-Tyrosine solution <![CDATA[A1]]> 50 μL 25 μL 25 μL 100 μL <![CDATA[A2]]> 75 μL 25 μL - 100 μL <![CDATA[A3]]> 50 μL 25 μL 25 μL 100 μL <![CDATA[A4]]> 75 μL 25 μL - 100 μL

[0105] Using a 96-well plate, add 25 μL of the sample solution and 25 μL of the tyrosinase solution, then add 100 μL of L-DOPA solution to initiate the reaction. Incubate with shaking at 37 °C in an enzyme-linked immunosorbent assay (ELISA) reader for 30 min, and scan and detect at a wavelength of 475 nm, and record the data.

[0106] For the absorbance of different extract samples, using vitamin C as the positive control, set three wells for each group and take the average value. The calculation formula for the inhibition rate is as follows:

[0107] Inhibition rate (%) = [1 - (A1 - A2) / (A3 - A0)] × 100%,

[0108] Where: A1 is the sample group, A2 is the control group of the sample group, A3 is the blank group, and A0 is the blank control group.

[0109] (6) The chemical components of the active compounds of Lycopodiastrum cernuum prepared in the examples and the IC 50 value of the tyrosinase activity inhibition ability of the control group vitamin C are shown in Table 2.

[0110] Table 2 Inhibition ability against tyrosinase

[0111] Group <![CDATA[IC 50 (mM)]]> Example 1 0.988 Example 2 1.350 Example 3 3.690 Example 4 7.959 Example 5 6.469 Example 6 3.216 Example 7 3.324 Example 8 2.437 Control group 0.984

[0112] As can be seen from Table 2, the active compounds I-VIII prepared in the examples all have inhibitory effects on tyrosinase. Among them, the active compound I provided in Example 1 exhibits inhibitory activity comparable to that of the positive control, and the IC 50 value is 0.988 mM. The active compounds II-VIII prepared in Examples 2-8 have good inhibitory activities, and the IC 50 values are 1.350 mM, 3.690 mM, 7.959 mM, 6.469 mM, 3.216 mM, 3.324 mM, and 2.437 mM, respectively.

[0113] The results show that the active compounds of Lycopodiastrum cernuum prepared in the examples all have high potential in the development of whitening products.

[0114] Therefore, the present invention adopts the above-mentioned active compound of Lycopodiastrum cernuum, its preparation method and application, discovers a new compound, whose tyrosinase activity is equivalent to that of the positive control, and solves the technical problems that existing whitening substances have risks such as allergy and carcinogenesis, and are prone to discoloration under the influence of the environment, thus affecting the whitening effect; the active compound of Lycopodiastrum cernuum with good whitening effect is obtained by a simple extraction method.

[0115] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention and are not intended to limit them. Although the present invention has been described in detail with reference to the preferred embodiments, those of ordinary skill in the art should understand that they can still modify or equivalently replace the technical solutions of the present invention, and these modifications or equivalent replacements cannot make the modified technical solutions deviate from the spirit and scope of the technical solutions of the present invention.

Claims

1. A Lycopodiella cuneata active compound, characterized in that: The structure of the compound is: ； Wherein, I is farlinol.

2. A method for preparing the compound as claimed in claim 1, comprising the following steps: Step S1, naturally air-drying and crushing the Lycopodiella ciliata sample to obtain powder; Step S2, soaking the powder obtained in step S1 with anhydrous ethanol, and concentrating under reduced pressure to obtain an ethanol extract; Step S3, dispersing the ethanol extract obtained in step S2, and extracting with petroleum ether and chloroform in sequence to obtain petroleum ether extract and chloroform extract; Step S4, separating the petroleum ether extract or chloroform extract obtained in step S3 on a 300-400 mesh silica gel chromatography column, eluting with petroleum ether-ethyl acetate mobile phase to obtain multiple components; further separating and purifying, and obtaining the active compounds of Lycopodiella foetida through an elution procedure.

3. The preparation method according to claim 2, characterized in that: In step S2, the powder obtained in step S1 is soaked in anhydrous ethanol for 3 times, each time for 7 days; the volume ratio of anhydrous ethanol to powder is 1:

1.

4. The preparation method according to claim 3, characterized in that: In step S2, the temperature of reduced pressure concentration is 37°C, the vacuum degree is 0.1 MPa, the rotation speed is 70 rpm / min, and the condensed water temperature is 5°C.

5. The preparation method according to claim 2, characterized in that: In step S4, the silica gel chromatography column is one or both of a 300-400 mesh silica gel chromatography column or a Sephadex LH-20 gel column chromatography column.

6. The preparation method according to claim 5, characterized in that: In step S4, the mobile phase in the elution procedure is one or more of petroleum ether-ethyl acetate, dichloromethane-methanol, and petroleum ether-dichloromethane-methanol.

7. Use of the compound according to claim 1 in the preparation of tyrosinase inhibitors.

8. Use of the compound according to claim 1 in preparing skin care products.

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