Marine fungus-derived super-sulfur compound as well as preparation method and application thereof

Abstract

The invention belongs to the technical field of biological medicines, and particularly relates to a marine fungus-derived super-sulfur compound as well as a preparation method and application thereof. The invention provides a super-sulfur compound which can promote proliferation and migration of human skin keratinocytes, has the potential of repairing skin injury and can be used for preparing a medicine for repairing skin injury. Moreover, the compound can be obtained by utilizing a sea calamus stem-derived fungus through a modern microbial fermentation engineering technology, large-scale industrial production is easy to realize, and the compound has wide application potential and development value in the field of skin repair medicines.

Description

Technical Field

[0001] This invention belongs to the field of biomedical technology. More specifically, it relates to a class of supersulfur compounds derived from marine fungi, their preparation methods, and applications. Background Technology

[0002] Sulfur, an essential element for the human body, plays a vital role in maintaining healthy hair, nails, and skin. In recent years, "supersulfides," composed of multiple sulfur atoms held together by elements such as carbon at both ends, have attracted much attention due to their unique biological activities. Studies have shown that these molecules not only have stronger antioxidant effects than vitamin E, but also exhibit various physiological functions such as anti-inflammatory and antiviral activity (Tsutsuki H, Zhang T, Akaike T, Sawa T. Regulation of innate immune and inflammatory responses by supersulfides. Int Immunol. 2024; 36(4):143-154). Of particular note is the significant effect of supersulfides on joint homeostasis and bone regeneration (Maemura M, Morita M, Ogata S, et al. Supersulfides contribute to joint homeostasis and bone regeneration. Redox Biol. 2025; 81:103545.), suggesting their important potential in the field of tissue repair.

[0003] Currently, commonly used skin damage repair drugs, such as recombinant human epidermal growth factor (EGF gel) and basic fibroblast growth factor (bFGF), need to be prepared through heterologous expression, which leads to problems such as high production costs, high technical difficulty, and unstable activity.

[0004] Therefore, the development of novel and highly effective tissue repair drugs has become an urgent need in clinical treatment, and supersulfides may provide a new solution for this due to their unique biological properties. Summary of the Invention

[0005] The technical problem to be solved by the present invention is to overcome the defects and shortcomings of existing skin damage repair drugs that rely on heterologous expression for preparation, and to provide a class of supersulfur compounds.

[0006] The purpose of this invention is to provide a method for preparing the supersulfur compound.

[0007] Another object of the present invention is to provide applications of the supersulfur compound.

[0008] Another objective of this invention is to provide a skin damage repair drug.

[0009] The above-mentioned objective of this invention is achieved through the following technical solution:

[0010] This invention protects a class of supersulfur compounds, characterized in that the supersulfur compounds have the structure shown in formula (I) or (II):

[0011]

[0012] Furthermore, the supersulfur compound is replaced with its pharmaceutically acceptable salt or solvate thereof.

[0013] The present invention also protects a method for preparing the supersulfur compound, wherein the supersulfur compound is obtained by extraction, separation and purification of fermentation products of fungi derived from the stem of *Acorus calamus*.

[0014] The fungus derived from the stem of the sea calamus is named Aspergillus fumigatiaffinis SYSU-6786, and was deposited at the Guangdong Provincial Center for Microbial Culture Collection on June 18, 2025, with accession number GDMCC No: 66541.

[0015] Furthermore, the preparation method specifically includes the following steps:

[0016] S1. The fungus Aspergillus fumigatiaffinis SYSU-6786 was activated and cultured on a large scale to obtain the fermentation product;

[0017] S2. Soak the fermentation product obtained in step S1 in methanol, filter, remove the solvent from the filtrate to obtain crude extract, add ethyl acetate to extract, remove the solvent from the upper organic layer to obtain extract.

[0018] S3. The extract obtained in step S2 is subjected to silica gel column adsorption, and gradient elution is performed using methanol-water solution as the eluent. The fraction with a methanol-water volume ratio of 30:70 is collected.

[0019] S4. The fraction with a methanol-water volume ratio of 30:70 obtained in step S3 is separated and purified by high performance liquid chromatography to obtain the supersulfur compound.

[0020] Further, in step S3, the gradient elution is performed using methanol-water solution as the eluent, with the methanol-water volume ratios being 10:90, 20:80, 30:70, 40:60, 50:50, 60:40, 70:30, 80:20, 90:10, and 100:0.

[0021] Furthermore, the conditions for high performance liquid chromatography in step S4 are as follows: the mobile phase is acetonitrile-water solution containing 0.1 vol% acetic acid, the flow rate is 3 mL / min, the detection wavelength is 254 nm, and the retention time is 0–40 min.

[0022] Further, the volume ratio of the acetonitrile to the aqueous solution containing 0.1 vol% acetic acid is (15-45):(85-55).

[0023] Specifically, the acetonitrile content in the mobile phase increases uniformly, with the volume percentage of acetonitrile gradually increasing from 15% to 45% within 40 minutes.

[0024] Furthermore, according to the method of the present invention, the supersulfur compounds can be distinguished by the elution order of the chromatographic peaks, wherein supersulfur compound (I) has a shorter retention time and elutes before supersulfur compound (II). Within a defined retention time range, the components corresponding to each chromatographic peak can be collected separately, and the structures of the obtained components can then be confirmed by nuclear magnetic resonance (NMR) technology.

[0025] Further, in step S3, the silica column is a reversed-phase silica column, specifically a Purospher STAR RP-18 column, 10×30cm, 5μm.

[0026] Furthermore, the chromatographic column used in the high-performance liquid chromatography is a semi-preparative column, specifically the Ultimate XB-C18 semi-preparative column, 10×250mm, 5μm; the instrument is an Essentia LC-16.

[0027] Furthermore, in step S1, the activation and expansion culture involves first preparing a seed culture solution and then carrying out fermentation culture.

[0028] Furthermore, the seed culture medium used in the preparation of the seed culture solution is PDB medium.

[0029] Preferably, the seed culture solution is prepared using a culture medium containing 25–35 g / L sea salt and 20–30 g / L PDB culture medium powder.

[0030] Furthermore, the PDB culture medium is a commercially available PDB culture medium.

[0031] Preferably, the seed culture medium is cultured under the following conditions: shaking speed of 100-200 rpm (preferably 150-180 rpm) and constant temperature culture at 25-30℃ for 3-5 days.

[0032] Furthermore, the culture medium used for the fermentation culture is rice culture medium.

[0033] Furthermore, the rice culture medium is prepared by mixing 40-50g of rice with 50mL of 2wt%-3wt% seawater. When needed, it can be sterilized at high temperature according to the above ratio.

[0034] Furthermore, the fermentation culture conditions are static culture at room temperature for 30 to 40 days.

[0035] Furthermore, the solvent removal is achieved by reducing pressure and concentrating the solvent.

[0036] This invention protects the use of the supersulfur compound in the preparation of skin damage repair drugs.

[0037] Preferably, the concentration of the supersulfur compound used on cells is 3–30 μmol / L (μM), more preferably 3.125–25 μM.

[0038] Furthermore, when the supersulfur compound has the structure shown in formula (I), the concentration of the supersulfur compound used on cells is 6 to 15 μM, preferably 6.25 to 12.5 μM.

[0039] Furthermore, when the supersulfur compound has the structure shown in formula (II), the concentration of the supersulfur compound used on cells is 3 to 30 μM, preferably 3.125 to 25 μM, and more preferably 6.25 to 12.5 μM.

[0040] Furthermore, the skin injury includes physical or chemical injury, specifically mechanical injury (such as abrasions, cuts, contusions, fractures, lacerations, punctures, gunshot wounds), temperature-related injury (burns, frostbite), electrical injury, corrosive injury, skin ulcers, etc.

[0041] Furthermore, the repair is to promote the repair and / or regeneration of skin wounds or scars.

[0042] Specifically, the repair promotes the migration and proliferation of HaCaT cells in the keratinocytes of the skin.

[0043] This invention also protects a skin damage repair drug, which includes one or two of the supersulfur compounds.

[0044] Compared with the prior art, the present invention has the following beneficial effects:

[0045] This invention provides a class of supersulfur compounds that can promote the proliferation and migration of human skin keratinocytes, possessing the potential to repair skin damage and can be used to prepare skin damage repair drugs. Furthermore, these compounds can be obtained using fungi derived from the stems of *Acorus calamus* through modern microbial fermentation engineering technology, facilitating large-scale industrial production and demonstrating broad application potential and development value in the field of skin repair drugs. Attached Figure Description

[0046] Figure 1 This is a plate colony diagram of fungi derived from the stem of *Acorus calamus* in Example 1 of the present invention.

[0047] Figure 2 This is a microscopic morphological image of a plate colony of Aspergillus fumigatiaffinis SYSU-6786, a fungus derived from the stem of Acorus calamus in Example 1 of this invention.

[0048] Figure 3 This is the carbon NMR spectrum of the supersulfur compound (I) obtained in Example 2 of the present invention.

[0049] Figure 4 This is the 1H NMR spectrum of the supersulfur compound (I) obtained in Example 2 of the present invention.

[0050] Figure 5 This is the carbon NMR spectrum of the supersulfur compound (II) obtained in Example 2 of the present invention.

[0051] Figure 6 This is the 1H NMR spectrum of the supersulfur compound (II) obtained in Example 2 of the present invention.

[0052] Figure 7 This is a statistical diagram (LSD, Mean±SD) of the cell scratch area after the supersulfur compound (I) obtained in Example 3 of the present invention was applied to human skin keratinocytes HaCaT cells.

[0053] Figure 8 This is a statistical diagram (LSD, Mean±SD) of the cell scratch area after the supersulfur compound (II) obtained in Example 3 of the present invention was applied to human skin keratinocytes HaCaT cells.

[0054] Figure 9 This is a diagram showing the changes in cell scratches after the supersulfur compound (I) obtained in Example 3 of this invention was applied to human skin keratinocytes (HaCaT cells).

[0055] Figure 10 This is a diagram showing the changes in cell scratches after the supersulfur compound (II) obtained in Example 3 of this invention was applied to human skin keratinocytes (HaCaT cells).

[0056] Figure 11The above is a statistical graph (K test, Mean±SD) of the cell activity of human skin keratinocytes after the supersulfur compounds (I) and (II) obtained in Example 3 of this invention were applied. Compared with the cn group, * indicates significant upregulation and # indicates significant downregulation (p<0.05). Detailed Implementation

[0057] The present invention will be further described below with reference to the accompanying drawings and specific embodiments, but the embodiments do not limit the present invention in any way. Unless otherwise specified, the reagents, methods and equipment used in the present invention are conventional reagents, methods and equipment in this technical field.

[0058] Unless otherwise specified, all reagents and materials used in the following examples are commercially available.

[0059] PDB medium (short for Potato Dextrose Broth), commercially available.

[0060] PDA medium (short for Potato Dextrose Agar (Medium)) was purchased commercially and prepared according to the instructions. It was then sterilized in an autoclave at 121°C (0.1 MPa) for 25 minutes. Before it solidified, the plates were poured out and cooled to room temperature to obtain PDA culture plates for later use.

[0061] Seed culture medium: Prepared by adding 30g of sea salt and 24g of PDB culture medium powder per liter of water, sterilized in an autoclave at 121℃ (0.1MPa) for 25 minutes, and cooled to room temperature for later use.

[0062] The fermentation medium is rice culture medium, with the following composition: 50g rice, 50mL 3% sea salt water, mixed in a culture bottle, sealed, and sterilized at 121℃ (0.1MPa) for 25min in a high-temperature sterilizer. It is then cooled to room temperature for later use.

[0063] Example 1: Obtaining Aspergillus fumigatiaffinis SYSU-6786, a fungus derived from the stem of Acorus calamus.

[0064] The supersulfur compounds (I) and (II) of this invention can be isolated and purified from the fermentation broth of the fungus strain Aspergillus fumigatiaffinis SYSU-6786, which originates from the stem of *Acorus calamus*. The specific isolation, purification, and identification steps for the fungus Aspergillus fumigatiaffinis SYSU-6786 are as follows:

[0065] 1. Isolation of Aspergillus fumigatiaffinis SYSU-6786 from the stem of Acorus calamus: The surface of the Acorus calamus stem was washed with sterile water, then the stem was chopped and inoculated into PDA culture plates. The plates were incubated at 25°C for 3-5 days. Colonies with different morphologies were picked and transferred to new PDA culture plates. The operation was repeated until a single morphological colony was obtained. The culture medium with a single colony was sent to Beijing Qingke Biotechnology Co., Ltd. for identification and ITS sequencing.

[0066] 2. Identification of Aspergillus fumigatiaffinis SYSU-6786

[0067] (1) Culture characteristics, microscopic examination and morphological features

[0068] The optimal culture temperature for this strain is 25–30℃, with humidity maintained at around 80%. Its morphology is fluffy or somewhat fibrous; upon maturity, it darkens in color, produces spores, and the colonies turn bluish-gray. The conidia have columnar heads and flask-shaped apical vesicles (e.g., Figures 1-2 (As shown).

[0069] (2) ITS gene sequence identification

[0070] The ITS-rRNA gene fragment sequence of this strain was obtained by sequencing, as shown in SEQ ID NO: 1, and is as follows:

[0071] .

[0072] After molecular biological identification, it was classified as Aspergillus fumigatiaffinis SYSU-6786 and deposited on June 18, 2025 at the Guangdong Provincial Microbial Culture Collection Center (GDMCC), located at No. 100, Xianlie Middle Road, Yuexiu District, Guangzhou, Guangdong Province, with accession number GDMCC No: 66541.

[0073] Example 2 Extraction and characterization of supersulfur compounds (I) and (II)

[0074] The supersulfur compounds (I) and (II) of this embodiment can be isolated and purified from the fermentation broth of the fungus strain Aspergillus fumigatiaffinis SYSU-6786, which originates from the stem of Acorus calamus. The specific process is as follows:

[0075] S1. Inoculate the fungus Aspergillus fumigatiaffinis SYSU-6786 into the seed culture medium and culture it on a shaker (25℃, 150-180rpm) to obtain the seed culture solution;

[0076] S2. Inoculate the seed culture medium obtained in step S1 into the fermentation medium and culture it at room temperature for 5 weeks (i.e. 35 days) to obtain the fermentation product.

[0077] S3. Soak the fermentation product obtained in step S2 in methanol for 3 days, filter, take the filtrate and concentrate under reduced pressure and extract with ethyl acetate, take the upper organic layer and concentrate under reduced pressure to obtain crude extract.

[0078] S4. The crude extract obtained in step S3 was adsorbed onto a silica gel column (specifically a Purospher STAR RP-18 reverse silica gel column, 10×30cm) and sequentially eluted with methanol-water solutions of volume ratios of 10:90, 20:80, 30:70, 40:60, 50:50, 60:40, 70:30, 80:20, 90:10, and 100:0. The fraction with a methanol-water volume ratio of 30:70 was collected.

[0079] S5. The methanol-water fraction obtained in step S3 with a volume ratio of 30:70 is separated and purified by high-performance liquid chromatography (HPLC). The HPLC conditions are as follows: mobile phase: acetonitrile-water solution containing 0.1 vol% acetic acid (the volume ratio of acetonitrile to the water solution containing 0.1 vol% acetic acid is (15-45):(85-55), and the acetonitrile content increases uniformly within 0-40 min); column: semi-preparative column Ultimate XB-C18, 10×250 mm, 5 μm; instrument: Essentia LC-16; flow rate: 3 mL / min; detection wavelength: 254 nm; components with retention times of 0-40 min are collected. The first peak appearing within the provided retention time (0-40 min) is supersulfur compound (I), and the third peak is supersulfur compound (II). That is, this invention distinguishes the supersulfur compounds by the elution order of the chromatographic peaks, wherein supersulfur compound (I) has a shorter retention time and elutes before supersulfur compound (II). Within a defined retention time range, the components corresponding to each chromatographic peak can be collected separately, and the structures of the obtained components can then be confirmed by nuclear magnetic resonance (NMR) technology.

[0080] The obtained supersulfur compound (I) was a yellow solid. Nuclear magnetic resonance (NMR) analysis of it yielded the following results: Figures 3-4 As shown, the physicochemical properties of this compound, as determined by analysis and testing, are as follows:

[0081] NMR data: 13C NMR(150MHz,DMSO-d6)δ:35.7,55.7,59.5,60.3,61.3,64.9,71.8,73.6,76.4 ,86.5,103.0,116.5,125.6,128.5,129.8,135.7,147.7,153.1,162.4,163.4.

[0082] 1 H NMR (600MHz, DMSO-d6) δ: 9.46 (s, 1H), 9.28 (d, J = 3.2Hz, 1H), 7.62 (d, J = 9.0Hz, 1H), 6.52 (d ,J=9.0Hz,1H),5.50(dt,J=2.5,10.3Hz,1H),5.43(d,J=10.3Hz,1H),5.29(d,J=5.2Hz,1H) ,5.28(s,1H),5.08(d,J=6.6Hz,1H),4.69(d,J=3.2Hz,1H),4.33(m,1H),4.20(m,1H),4.01 (d,J=7.1Hz,1H),3.78(s,3H),3.67(s,3H),2.43(d,J=15.2Hz,1H),1.96(d,J=15.2Hz,1H).

[0083] Analysis of the nuclear magnetic resonance (NMR) results confirmed the molecular formula of the obtained compound to be: C 20 H 22 N2O9S3 has the structure of formula (I):

[0084]

[0085] The obtained supersulfur compound (II) was a light yellow solid. Nuclear magnetic resonance (NMR) analysis of it yielded the following spectrum: Figures 5-6 As shown, the physicochemical properties of this compound, as determined by analysis and testing, are as follows:

[0086] NMR data: 13 C NMR(150MHz,DMSO-d6)δ:35.8,56.2,60.3,64.8,69.4,71.6,73.4,76.5,86.9 ,97.7,105.8,117.7,118.2,128.6,129.6,132.7,150.1,153.9,161.3,163.2.

[0087] 1H NMR(600MHz,DMSO-d6)δ:10.35(s,1H),6.73(d,J=8.2Hz,1H),6.59(d,J=8.2Hz ),5.50(s,1H),5.49(dt,J=10.4,2.5Hz,1H),5.43(d,J=10.4Hz,1H),5.33(d,J =5.4Hz,1H),5.29(d,J=6.6Hz,1H),4.29(m,1H),4.19(m,1H),4.03(d,J=7.1Hz ,1H),3.77(s,3H),3.73(s,3H),2.44(d,J=15.2Hz,1H),2.04(d,J=15.2Hz,1H).

[0088] Analysis of the nuclear magnetic resonance (NMR) results confirmed the molecular formula of the obtained compound to be: C 20 H 20 N2O9S3 has the structure of formula (II):

[0089]

[0090] Example 3: Activity Test of Supersulfur Compounds

[0091] 1. Experimental materials:

[0092] 12-well plates for counting, trypsin, double antibiotics (streptomycin / penicillin), fetal bovine serum, incubator, PBS buffer, cell counting chamber.

[0093] 2. Experimental methods:

[0094] (1) Cell culture: HaCaT cells were cultured in DEME medium containing double antibiotics (streptomycin / penicillin, 1%) and fetal bovine serum (10%). The culture conditions were 37°C and 5% CO2 constant temperature incubator.

[0095] (2) Cell passage: When the cell density reaches 80% to 100%, remove the culture medium, wash gently with preheated PBS, add 2 mL of 0.05% trypsin to digest for 2 to 5 minutes until the cell edges become rounded or partially and completely detach from the culture dish, add 5 mL of fresh complete culture medium to stop digestion, centrifuge at 1000 rpm for 2 minutes, discard the supernatant, add fresh complete culture medium to resuspend and passage culture at an appropriate ratio.

[0096] (3) Scratch test

[0097] DMSO was used as a solvent to prepare 10 mM mother liquors of supersulfur compound (I) and supersulfur compound (II).

[0098] Cells were seeded at a density of 15 mg / well in 12-well plates and incubated overnight in a 5% CO2 incubator at 37°C. The next day, cells were streaked, and each well was gently washed with 500 μL of pre-warmed PBS, followed by photographic recording. Using DMEM medium, 10 mM stock solutions of supersulfur compound (I) and supersulfur compound (II) were diluted to final concentrations of 3.125 μM, 6.25 μM, 12.5 μM, 25 μM, and 50 μM, and added to the 12-well plates, with photographic recording. After 48 h of incubation, the plates were photographed again.

[0099] (4) MTT cell viability / toxicity assay

[0100] Cells were seeded at a density of 5000 cells / well in 96-well plates (margin wells were filled with sterile PBS) and incubated overnight in a 5% CO2, 37°C incubator. The next day, the culture medium was discarded, and the cells were carefully rinsed 1-2 times with PBS. 100 μL of a compound diluted with DMEM medium was added to each experimental well (the compound was diluted with DMSO at a stock concentration of 10 mM, ensuring a final DMSO concentration <1%). Only 100 μL of DMEM medium was added to each negative control well (this was the untreated group, or cn group). Cells were incubated at this temperature for 48 hours, then culture medium containing 1% MTT (5 mg / mL) was added. After another 4 hours of incubation in the dark, the culture medium was carefully aspirated from each well, and 150 μL of DMSO was added to each well. The plates were then shaken at 300 rpm for 10 minutes, and the absorbance was measured at 570 nm. Cell viability was calculated as [(experimental wells - blank wells) / (negative control wells - blank wells)] × 100%. The wells include negative control wells (containing cell-containing culture medium and MTT) and blank wells (containing cell-free and test-substance-free culture medium, MTT, and DMSO).

[0101] 3. Experimental Results:

[0102] Cell scratch assay results are as follows Figures 7-10 As shown, two days after administration, the final concentrations of supersulfur compound (I) at 6.25 μM and 12.5 μM promoted the proliferation and migration of human keratinocytes (HaCaT cells), and the final concentrations of supersulfur compound (II) at 6.25 μM, 12.5 μM and 25 μM also promoted the proliferation and migration of human keratinocytes (HaCaT cells).

[0103] Depend on Figure 11 It is evident that supersulfur compound (I) at a final concentration ≤12.5 μM and supersulfur compound (II) at a final concentration ≤50 μM did not exhibit cytotoxicity against human keratinocyte HaCaT cells. Furthermore, supersulfur compound (II) significantly improved the survival rate of HaCaT cells at concentrations of 3.125 μM, 6.25 μM, and 12.5 μM.

[0104] In summary, the supersulfur compounds (I) and (II) of this invention have a certain effect on promoting the proliferation and migration of human skin keratinocytes (HaCaT cells), and can be considered as lead compounds for subsequent drug development at appropriate concentrations.

[0105] The above embodiments are preferred embodiments of the present invention, but the embodiments of the present invention are not limited to the above embodiments. Any changes, modifications, substitutions, combinations, or simplifications made without departing from the spirit and principle of the present invention shall be considered equivalent substitutions and shall be included within the protection scope of the present invention.

Claims

1. A class of supersulfur compounds, characterized in that, The supersulfur compound has the structure shown in formula (I) or (II):

2. The supersulfur compound according to claim 1, characterized in that, The supersulfur compound is replaced with its pharmaceutically acceptable salt or its solvate.

3. The method for preparing the supersulfur compound according to claim 1, characterized in that, The supersulfur compound was obtained by extraction, separation, and purification of the fermentation products of fungi derived from the stems of *Acorus calamus*. The fungus derived from the stem of the sea calamus is named Aspergillus fumigatiaffinis SYSU-6786, and was deposited at the Guangdong Provincial Center for Microbial Culture Collection on June 18, 2025, with accession number GDMCC No: 66541.

4. The preparation method according to claim 3, characterized in that, The preparation method specifically includes the following steps: S1. The fungus Aspergillus fumigatiaffinis SYSU-6786 was activated and cultured on a large scale to obtain the fermentation product; S2. Soak the fermentation product obtained in step S1 in methanol solvent, filter, remove the solvent from the filtrate to obtain crude extract, add ethyl acetate to extract, remove the solvent from the upper organic layer to obtain extract. S3. The extract obtained in step S2 is subjected to silica gel column adsorption, and gradient elution is performed using methanol-water solution as the eluent. The fraction with a methanol-water volume ratio of 30:70 is collected. S4. The fraction with a methanol-water volume ratio of 30:70 obtained in step S3 is separated and purified by high performance liquid chromatography to obtain the supersulfur compound.

5. The preparation method according to claim 4, characterized in that, In step S3, the gradient elution is performed using methanol-water solution as the eluent, with the methanol-water volume ratios being 10:90, 20:80, 30:70, 40:60, 50:50, 60:40, 70:30, 80:20, 90:10, and 100:

0.

6. The preparation method according to claim 4, characterized in that, The conditions for high performance liquid chromatography in step S4 are as follows: the mobile phase is acetonitrile-water solution containing 0.1 vol% acetic acid, the flow rate is 3 mL / min, the detection wavelength is 254 nm, and the retention time is 0–40 min.

7. The preparation method according to claim 6, characterized in that, The volume ratio of the acetonitrile to the aqueous solution containing 0.1 vol% acetic acid is (15-45):(85-55).

8. The preparation method according to claim 4, characterized in that, The high-performance liquid chromatography column used is a semi-preparative column.

9. The use of the supersulfur compound of claim 1 or 2 in the preparation of a skin damage repair drug.

10. A skin damage repair drug, characterized in that, The skin damage repair drug includes one or two of the supersulfur compounds described in claim 1 or 2.

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