PRDX1 inducer and use thereof in preparing medicament against acute kidney injury

Abstract

The present application relates to the field of natural products, and specifically, to a PRDX1 inducer. The PRDX1 inducer comprises 13-hydroxyglucopiericidin A, a piericidin glucoside, represented by formula (I), or a pharmaceutically acceptable salt thereof. The PRDX1 inducer can be used to prepare a medicament against acute kidney injury with improved therapeutic effects in preventing and treating an acute kidney disease.

Description

A PRDX1 inducer and its application in preparing anti-acute kidney injury drugs Technical Field

[0001] The present invention belongs to the field of natural products, and specifically relates to a PRDX1 inducer and an application thereof in the preparation of an anti-acute kidney injury drug. Background Art

[0002] Acute kidney injury (AKI) is a clinical syndrome characterized by a rapid decline in renal function, leading to elevated serum creatinine levels and / or decreased urine output. Its occurrence is closely related to sepsis, renal ischemia-reperfusion injury, and nephrotoxic drugs. To date, the pathophysiology of AKI has not been fully elucidated, but damage and death of renal tubular cells (especially proximal tubular cells) play a key role. However, severe or persistent damage often leads to insufficient and incomplete repair, resulting in tubular degeneration, inflammation, renal fibrosis, and ultimately progression to chronic kidney disease (CKD) or end-stage renal disease. Currently, in addition to supportive care, there is no effective and targeted treatment for AKI in clinical practice. Therefore, effective treatment strategies based on the pathogenesis of AKI are urgently needed.

[0003] Summary of the Invention

[0004] In order to solve the problem that the treatment effect of acute kidney injury in the prior art is unsatisfactory, the present invention provides a PRDX1 inducer.

[0005] The PRDX1 inducer comprises 13-hydroxyglucopiericidin A or a pharmaceutically acceptable salt thereof,

[0006] Furthermore, the pieridius mycin glucoside 13-hydroxyglucopiericidin A or a pharmaceutically acceptable salt thereof inhibits the generation of ROS by activating the PRDX1 / Nrf2 pathway.

[0007] Furthermore, the pieridial mycin glucoside 13-hydroxyglucopiericidin A or a pharmaceutically acceptable salt thereof binds to Cys83 of the PRDX1 and increases the activity of the peroxidase PRDX1, thereby inducing the PRDX1 to enter the nucleus, bind to the nuclear transcription factor Nrf2, and activate the Nrf2 / HO-1 / NQO1 pathway.

[0008] One object of the present invention is to provide use of the PRDX1 inducer described in any one of the above items in the preparation of an anti-acute kidney injury drug.

[0009] Furthermore, in the use of a PRDX1 inducer in the preparation of an anti-acute kidney injury drug, the PRDX1 inducer alleviates the symptoms of acute kidney injury by attenuating necrosis and shedding of renal tubular epithelial cells in mice with acute nephritis.

[0010] Furthermore, in the use of the PRDX1 inducer in the preparation of anti-acute kidney injury drugs, the PRDX1 inducer induces the binding of Cys83 of the PRDX1 and increases the activity of the peroxidase PRDX1, induces the PRDX1 to enter the nucleus, binds to the nuclear transcription factor Nrf2, activates the Nrf2 / HO-1 / NQO1 pathway, and inhibits the production of ROS.

[0011] Furthermore, the acute kidney injury is caused by ischemia-reperfusion.

[0012] The present invention also provides a method for preparing pieridialmycin glucoside 13-hydroxyglucopiericidin A.

[0013] The preparation method of pieridiomycin glucoside 13-hydroxyglucopiericidin A comprises the following steps:

[0014] The fermentation product of Streptomyces psammoticus SCSIO NS126 was extracted with ethyl acetate, concentrated, and eluted with a methanol / water gradient of 1:9, 3:7, 5:5, 7:3, and 10:0, respectively. The fractions eluted with a methanol / water volume ratio of 7:3 were purified to obtain 13-hydroxyglucopiericidin A.

[0015] Furthermore, the purification comprises the following steps:

[0016] The obtained fractions were purified by HPLC liquid chromatography, which included a 10 mm × 250 mm, 5 μm, YMC-pack ODS-A chromatographic column, a v / v mobile phase of acetonitrile:water = 45:55, and a flow rate of 2.5 mL / min.

[0017] Furthermore, the Streptomyces psammoticus SCSIO NS126 is cultured on a solid state and expanded to obtain a fermentation product. The culture medium used for the solid culture comprises 6 g of yeast extract powder, 10 g of malt extract powder, 12 g of glucose, 20 g of agar, and 1 L of water. The pH is adjusted to 7.2 and the culture medium is sterilized.

[0018] The culture medium for expanded culture is selected from a first culture medium or a second culture medium. The first culture medium includes 6 g of yeast extract powder, 10 g of malt extract powder, 12 g of glucose, 20 g of agar, and 1 L of water, and is adjusted to pH 7.2 and sterilized. The second culture medium includes 10 g of glucose, 3 g of yeast extract powder, 10 g of starch, 25 g of cottonseed powder, 2 g of NaCl, 5 g of CaCO3, and 1 L of water, and is adjusted to pH 4.0-7.2 and sterilized.

[0019] Compared with the prior art, the advantages of the present invention are:

[0020] 1. This invention confirms for the first time the PRDX1-inducing effect of compound S14, providing a new candidate compound for the preparation of drugs for the prevention and treatment of acute kidney injury.

[0021] 2. This invention is the first to explore the PRDX1 / Nrf2 signaling pathway in the occurrence and development of acute kidney injury, confirming that the PRDX1 / Nrf2 signaling pathway plays an important role in the occurrence and development of acute kidney injury. It is expected to be used in the development of more effective acute kidney injury therapeutic drugs targeting the PRDX1 / Nrf2 signaling pathway.

[0022] 3. The preparation method of pieridian mycin glucoside 13-hydroxyglucopiericidin A provided by the present invention can be used to prepare pieridian mycin glucoside 13-hydroxyglucopiericidin A in large quantities, thereby reducing production costs and shortening production time. BRIEF DESCRIPTION OF THE DRAWINGS

[0023] Figure 1. Mass spectrum of the product provided in Example 1;

[0024] Figure 2. Product provided in Example 1 1 H-NMR spectrum (DMSO-d6, 500 MHz);

[0025] Figure 3. Product provided in Example 1 13 C-NMR spectrum (DMSO-d6, 125 MHz);

[0026] FIG4 is a graph showing serum creatinine and urea nitrogen levels of S14 in UIRI mice provided in Example 2;

[0027] FIG5 is HE staining of S14 in UIRI mice provided in Example 2;

[0028] Figure 6 shows the ROS level and PRDX1 / Nrf2 pathway-related proteins (NQO1 and HO-1) of S14 in the H2O2-induced oxidative stress HK-2 cell model provided in Example 3;

[0029] FIG7 is an immunofluorescence image of S14-induced PRDX1 nuclear translocation provided in Example 3;

[0030] FIG8 is the immunoprecipitation and immunofluorescence detection of the interaction between PRDX1 and Nrf2 proteins provided in Example 3;

[0031] FIG9 is a graph showing the binding of S14 to PRDX1 protein detected by the SPR method and cellular thermal shift assay (CETSA)-western immunoblotting method provided in Example 3. DETAILED DESCRIPTION

[0032] In order to make the above-mentioned objects, features and advantages of the present invention more obvious and easy to understand, the specific embodiments of the present invention are described in detail below with reference to the accompanying drawings, but they should not be understood as limiting the scope of implementation of the present invention.

[0033] In various forms of AKI, harmful substances accumulate in the renal tubules, inducing immune cells, endothelial cells, and renal tubular endothelial cells to produce large amounts of reactive oxygen species (ROS), leading to a disruption of renal redox balance. Furthermore, renal hypoxia caused by persistent hypoperfusion, ischemia-reperfusion, or microcirculatory impairment leads to the generation of large amounts of ROS in mitochondria. Therefore, excessive ROS production is a central mechanism underlying oxidative stress, inflammation, and adaptive responses in AKI.

[0034] The peroxiredoxins (PRDXs) family is an important antioxidant that can effectively scavenge intracellular ROS and maintain biological stability. PRDX1 is a key member of this family. In the cytoplasm, PRDX1 has antioxidant functions.

[0035] The present invention discloses 13-hydroxyglucopiericidin A (S14), a pieritin glucoside, as a PRDX1 inducer for preventing and treating AKI by activating PRDX1. S14 primarily binds to Cys83 of PRDX1 and increases the activity of the peroxidase PRDX1. Furthermore, S14 increases PRDX1 nuclear translocation, allowing PRDX1 to bind to Nrf2 and activate the Nrf2 / HO-1 / NQO1 pathway, thereby inhibiting the production of reactive oxygen species (ROS). This invention provides a new lead compound for the development of drugs against acute kidney injury and is of great significance for the development of marine-derived drugs in China.

[0036] The molecular structure of 13-hydroxyglucopiericidin A is shown in the figure below:

[0037] The present invention obtains pieridialmycin glucoside S14 by performing shaker amplified fermentation and extraction and purification on a Streptomyces psammoticus SCSIO NS126 derived from mangrove sediment (deposited in Guangdong Provincial Microbial Culture Collection Center on April 19, 2024, with a deposit address of 5th Floor, Building 59, No. 100 Xianlie Middle Road, Guangzhou, with a deposit date of April 19, 2024 and a deposit number of GDMCC No. 64524).

[0038] Example 1:

[0039] The preparation method of pieridium glucoside S14 comprises the following steps:

[0040] 1. Solid Culture of Streptomyces psammoticus SCSIO NS126

[0041] Streptomyces psammoticus SCSIO NS126 (GDMCC No. 64524) was isolated from mangrove sediment and stored on a slant of ISP-2 culture medium. The main ingredients were: 6 g yeast extract powder, 10 g malt extract powder, 12 g glucose, 20 g agar, 1 L water, pH 7.2, and sterilized for later use.

[0042] 2. Expanded Fermentation of Streptomyces psammoticus SCSIO NS126

[0043] A small amount of the slant culture was used for seed liquid fermentation. The culture medium consisted of 6g yeast extract powder, 10g malt extract powder, 12g glucose, 20g agar, and 1L water, with a pH of 7.2, and was sterilized. The culture conditions were 28°C and 180rpm. After 48 hours of incubation, shaker fermentation was performed. The fermentation volume was 24L. The fermentation medium consisted of 20g soluble starch, 10g cottonseed meal, 5g yeast extract powder, 20g maltodextrin, 5g malt extract powder, 2g anhydrous magnesium sulfate, 2g sodium chloride, 2g calcium carbonate, and 1L water, with a pH of 7.0-7.2, and was sterilized. After 7 days of fermentation at 28°C and 180rpm, the fermentation broth was extracted with ethyl acetate, and the extract was concentrated to obtain a crude extract.

[0044] 3. Extraction and Separation

[0045] The crude extract obtained by fermentation of strain Streptomyces psammoticus SCSIO NS126 was dissolved in a small amount of methanol, and then mixed with ODS and subjected to reverse phase column (Spherical C18, 20-45 μm, ) Chromatography was performed with a methanol:water gradient elution (v / v, 1:9, 3:7, 5:5, 7:3, 10:0) to obtain 18 elution fractions B1 - B18. The fraction from the B12 fraction (eluted with v / v, methanol:water = 7:3) was purified by semi - preparative HPLC (Infinity 1260) using a YMC - pack ODS - A column, 10 mm × 250 mm, 5 μm, with a mobile phase of v / v, acetonitrile:water = 45:55 and a flow rate of 2.5 mL / min to obtain the product (6.9 mg, t R = 20.4 min).

[0046] The prepared product was analyzed by mass spectrometry, nuclear magnetic resonance 1H - NMR and 13C - NMR. Referring to Figure 1, the molecular ion peak of the product was 594.3273 [M + H] + , and its molecular weight was obtained as 593; referring to Figures 2 and 3, the 1H - NMR and 13C - NMR were consistent with the literature reports (J. Antibiot. 1990, 43, 1329), proving that the product prepared in Example 1 was piericidin glucoside S14.

[0047] Optionally, the medium in step 2 can be replaced with 10 g of glucose, 3 g of yeast extract powder, 10 g of starch, 25 g of cottonseed powder, 2 g of NaCl, 5 g of CaCO3, 1 L of water, adjusted to any value within pH 4.0 - 7.2, and sterilized for preparation. Fermentation was carried out in a fermenter. Other fermentation conditions remained unchanged.

[0048] Example 2:

[0049] This example investigated the effect of S14 on the renal function of UIRI mice

[0050] Experimental animals: Male C57BL / 6 mice, SPF grade, weighing 22 - 25 g, were provided by the Experimental Animal Center of Southern Medical University (License number: SCXK(Guangdong)2016 - 0041). All mice were housed in an environment with a temperature of 25 ± 1°C and a humidity of 55 ± 5%. The light - dark cycle was 12 h light and 12 h dark, and they had free access to water and food. They were housed in a SPF - level animal room for one week before the experiment to adapt to the environment.

[0051] 1. Compound S14 alleviates renal injury caused by UIRI

[0052] 1.1 Establishment of an acute kidney injury model induced by UIRI and grouped administration

[0053] Eighteen male C57 BL / 6 mice were randomly divided into three groups, with six mice in each group: a sham-operated (Sham) group, a UIRI model group, and a UIRI+S14 (1 mg / kg) group. The UIRI+S14 (1 mg / kg) group received an intraperitoneal injection of 1 mg / kg S14 solution once daily for three consecutive days. During this period, the sham-operated (Sham) and UIRI model groups received an equal dose of blank control solvent (0.1% DMSO / PBS, DMSO to PBS volume ratio of 1:1000) via intraperitoneal injection. On the third day, 4 hours after drug administration, mice in the UIRI model group and the UIRI+S14 (1 mg / kg) group underwent unilateral renal artery clamping for 45 minutes. The sham group only exposed the kidney without renal artery clamping. 48 hours later, blood was collected from the orbital venous plexus into a centrifuge tube containing sodium heparin and centrifuged at 8000 rpm for 5 minutes (4°C). The upper plasma layer was aspirated into a new centrifuge tube for the determination of plasma biochemical parameters. The mice were then sacrificed, and the kidney tissue was removed, a portion of which was fixed in 4% paraformaldehyde for histopathological examination.

[0054] 1.2 Serum creatinine and urea nitrogen levels

[0055] The serum creatinine and urea nitrogen test kits were developed in Nanjing, and the specific measurement methods were carried out in full accordance with the instructions.

[0056] 1.3 Observation of renal tissue pathology (HE staining)

[0057] The collected organ tissues were fixed in 4.5% formalin, embedded in paraffin, and sliced. The slices were stained with hematoxylin and eosin (H&E), and the morphological analysis of the tissues was performed by light microscopy.

[0058] 1.4 Experimental Results

[0059] Plasma biochemical indicators were detected using serum creatinine and urea nitrogen kits. As shown in Figure 4, the serum creatinine and urea nitrogen in the UIRI model group increased by 3.84 times and 3.25 times respectively compared with the Sham group, indicating that the unilateral renal artery clamping method was used to successfully induce the acute kidney injury model. The serum creatinine and urea nitrogen in the UIRI+S14 group were significantly reduced compared with the UIRI model group, indicating that compound S14 can alleviate the symptoms of acute nephritis and has a certain therapeutic effect on acute nephritis. As shown in Figure 5 (the triangles in the figure represent necrosis and shedding of renal tubular epithelial cells, the formation of cell fragments or cell casts in the lumen, and the ovals represent cell vacuolation), S14 significantly weakened the necrosis and shedding of renal tubular epithelial cells in mice induced by UIRI, and there were shedding fragments in the tubular lumen. The results show that S14 can alleviate the renal damage caused by UIRI by weakening the necrosis and shedding of renal tubular epithelial cells induced by UIRI in mice.

[0060] Example 3:

[0061] This example investigates the effect of compound S14 on reducing H2O2-induced apoptosis by regulating PRDX1 / Nrf2 in HK-2 (human renal tubular epithelial cells).

[0062] Experimental cells: HK-2 human renal tubular epithelial cells were cultured in DMEM / F12 medium containing 10% fetal bovine blood in a CO2 incubator (37°C, 5% CO2).

[0063] 1. Effects of compound S14 on ROS levels and PRDX1 / Nrf2 pathway-related proteins in H2O2-induced oxidative stress

[0064] 1.1 Cellular intervention

[0065] HK-2 cells cultured in vitro in the logarithmic growth phase were taken and 2×10 5 Cells were seeded at a density of 100 cells / well in a six-well plate and cultured for 24 hours. After the cells adhered, they were divided into four groups and co-cultured for 24 hours with the compound. The H2O2 stock solution was diluted with PBS to a concentration of 500mM. The S14 stock solution was dissolved in DMSO and the S14 stock solution concentrations were 5 and 10mM, respectively.

[0066] Control group: 2 uL DMSO was added to 2 mL of culture medium containing 10% FBS and co-cultured with cells.

[0067] H2O2 group: 2 uL of H2O2 stock solution and 2 uL of DMSO were added to 2 mL of culture medium containing 10% FBS and co-cultured with the cells.

[0068] H2O2+S14 (5 μM) group: 2 μL of H2O2 stock solution was added to 2 mL of culture medium containing 10% FBS, and co-cultured with cells for 4 h. Then, 2 μL of S14 stock solution (5 mM) was added for co-culture.

[0069] H2O2+S14 (10 μM) group: 2 μL of H2O2 stock solution was added to 2 mL of culture medium containing 10% FBS, and co-cultured with cells for 4 h. Then, 2 μL of S14 stock solution (10 mM) was added for co-culture.

[0070] 1.2 Detection of ROS and PRDX1 / Nrf2 pathway-related proteins (NQO1 and HO-1)

[0071] To detect the intracellular ROS level, DCFH-DA fluorescent probe (5 μM) was used to incubate four groups of HK-2 cells at 37°C in the dark for 30 min, the ROS working solution was discarded, the cells were washed three times with PBS, and the images were taken under an inverted fluorescence microscope.

[0072] To detect the protein expression levels of PRDX1 / Nrf2 and its downstream target genes NQO1 and HO-1, after the four groups of HK-2 cells were co-cultured for 24 h, the total protein in the HK-2 cells was extracted and the changes in protein expression were detected by Western Blot.

[0073] 1.3 Experimental Results

[0074] Images were taken using an inverted fluorescence microscope and then quantitatively analyzed. As shown in Figure 6, the H2O2 group produced a large amount of ROS, while the S14 group reduced the level of ROS in a concentration-dependent manner. Western blot was used to detect changes in the expression of proteins related to the PRDX1 / Nrf2 pathway. As shown in Figure 6, the protein expression of PRDX1 in the H2O2-treated group was 0.76 times that of the Control group. After S14 treatment, the PRDX1 protein expression levels were 1.41 times and 1.53 times that of the H2O2 group, respectively. The expression of Nrf2, NQO1, and HO-1 proteins also increased. The results show that S14 may significantly reduce ROS levels through the PRDX1 / Nrf2 pathway.

[0075] 2. S14 induces PRDX1 to enter the nucleus, where PRDX1 binds to the nuclear transcription factor Nrf2

[0076] 2.1 Take HK-2 cells cultured in vitro in the logarithmic growth phase and use 2×10 5 Cells were seeded at a density of 100 cells / well in a six-well plate and cultured for 24 hours. After the cells adhered to the wall, they were divided into three groups and the compound was added for 24 hours of co-culture. The S14 stock solution was dissolved in DMSO, and the concentrations of the S14 stock solution were 5 and 10 mM, respectively.

[0077] Control group: 2 μL of DMSO was added to 2 mL of culture medium containing 10% FBS and co-cultured with cells.

[0078] S14 (5 μM) group: 2 μL of S14 stock solution (5 mM) was added to 2 mL of culture medium containing 10% FBS and co-cultured with cells.

[0079] S14 (10 μM) group: 2 μL of S14 stock solution (10 mM) was added to 2 mL of culture medium containing 10% FBS and co-cultured with cells.

[0080] 2.2 Cell immunofluorescence

[0081] Cells were fixed with 4% paraformaldehyde and washed with graded alcohols. They were then blocked with blocking buffer (1% BSA and 0.5% Tween 20 in PBS) for 1 hour at room temperature. HK-2 cells were then incubated with primary antibodies overnight at 4°C. Cells were then incubated with Alexa Fluor 488 / 594-conjugated secondary antibodies for 2 hours at room temperature. Sections were then mounted with an anti-fluorescence quencher containing Hoechst 33342.

[0082] 2.3 Immunoprecipitation

[0083] For immunoprecipitation assays, plasmids Flag or Flag-PRDX1 were transfected into HK-2 cells according to the protocol provided by the Lipofectamine 3000 kit (Invitrogen, USA). 48 h after transfection, cells were harvested for Western blotting and immunoprecipitation analysis.

[0084] 2.4 Experimental Results

[0085] Immunofluorescence results showed that S14 promoted the translocation of PRDX1 and Nrf2 to the cell nucleus (Figures 7 (Nuclei showed blue fluorescence, PRDX1 showed green fluorescence, Merge showed blue and green fluorescence) and 8 (Nuclei showed blue fluorescence, PRDX1 showed red fluorescence, Nrf2 showed green fluorescence, Merge showed blue, red and green fluorescence)). To evaluate the binding between PRDX1 and Nrf2, immunoprecipitation and immunofluorescence showed that PRDX1 bound to Nrf2 (Figure 8). Some studies have shown that promoting the entry of Nrf2 can regulate the expression of downstream heme oxygenase-1 (HO-1), thereby inhibiting the production of ROS. These findings collectively indicate that PRDX1 binds to the nuclear transcription factor Nrf2 to regulate ROS levels.

[0086] Example 4:

[0087] This example investigates the binding of compound S14 to PRDX1 protein

[0088] 1. SPR analysis of the affinity of S14 for PRDX1 protein

[0089] 1.1 SPR method

[0090] SPR measurements were performed using the PlexArray HT A100 system (Plexera LLC, Bothell, WA, USA) to analyze the interaction. Human PRDX1 protein (Cloud Clone, RPC749Hu01, China) was immobilized on a nanocapture sensor chip with a bare gold coating. S14 was diluted in PBS buffer at concentrations of 0.3125, 0.625, 1.25, or 2.5 μM and injected in multi-cycle analysis mode. Data were analyzed using BIA evaluation software.

[0091] 1.2 Cellular Thermal Shift Assay (CETSA)

[0092] To evaluate the binding affinity of S14 to PRDX1, a cell thermal shift assay was performed. HK-2 cells were seeded in a large dish (1×10 6 cells), when the HK-2 cells reached ~70%, 1‰ DMSO or S14 (final concentration 10μM, final concentration adjusted with 1‰ DMSO) was added and incubated for 4 hours. Wash with PBS three times, then add PBS with protease inhibitors, scrape the cells with a scraper, and then transfer them to a cryopreservation tube. Use liquid nitrogen to freeze and thaw the cell suspension 5 times. Centrifuge the cell lysate at 13,000×g for 20 minutes at 4°C. Divide the respective supernatants into aliquots and heat them at different temperatures (45, 49, 53, 57, 61, 65°C) for 3 minutes respectively. Centrifuge the heated supernatant at 13,000×g for 15 minutes at 4°C to remove denatured proteins. Collect the supernatant and analyze it using Western blotting.

[0093] 1.3 Experimental Results

[0094] As shown in Figure 9 (the curves in the left figure represent 2.5μM, 1.25μM, 0.625μM, and 0.3125μM from top to bottom, respectively), SPR results showed that S14 bound to the PRDX1 protein in a concentration-dependent manner, with a dissociation equilibrium constant (KD) of 426nM. Next, the binding of PRDX1 and S14 was determined by CETSA-WB. The CETSA-WB results showed that compared with the DMSO group, S14 could significantly improve the thermal stability of PRDX1, further supporting the direct binding of S14 to PRDX1 as its target.

Claims

1. A PRDX1 inducer, characterized in that Including 13-hydroxyglucopiericidin A or a pharmaceutically acceptable salt thereof represented by formula (I), 2. The PRDX1 inducer according to claim 1, wherein The pieridius mycin glucoside 13-hydroxyglucopiericidin A or a pharmaceutically acceptable salt thereof inhibits the generation of ROS by activating the PRDX1 / Nrf2 pathway.

3. The PRDX1 inducer according to claim 2, wherein The pieridian mycin glucoside 13-hydroxyglucopiericidin A or a pharmaceutically acceptable salt thereof activates the PRDX1 / Nrf2 pathway to inhibit the production of ROS, including: the pieridian mycin glucoside S14 or a pharmaceutically acceptable salt thereof binds to Cys83 of the PRDX1 and increases the activity of the peroxidase PRDX1, inducing the PRDX1 to enter the nucleus, bind to the nuclear transcription factor Nrf2, and activate the Nrf2 / HO-1 / NQO1 pathway.

4. Use of the PRDX1 inducer according to any one of claims 1 to 3 in the preparation of an anti-acute kidney injury drug.

5. The use according to claim 4, characterized in that The PRDX1 inducer alleviates the symptoms of acute kidney injury by reducing the necrosis and shedding of renal tubular epithelial cells in mice with acute nephritis.

6. [Corrected 12.09.2024 under Rule 26] The application according to claim 4, characterized in that The PRDX1 inducer induces the Cys83 binding of the PRDX1 and increases the activity of the peroxidase PRDX1, induces the PRDX1 to enter the nucleus, binds to the nuclear transcription factor Nrf2, activates the Nrf2 / HO-1 / NQO1 pathway, and inhibits the generation of ROS.

7. [Corrected 12.09.2024 according to Rule 26] A method for preparing 13-hydroxyglucopiericidin A, characterized in that: The following steps are involved: The fermentation product of Streptomyces psammoticus SCSIO NS126 was extracted with ethyl acetate, concentrated, and eluted with a methanol / water gradient of 1:9, 3:7, 5:5, 7:3, and 10:0, respectively. The fractions eluted with a methanol / water volume ratio of 7:3 were purified to obtain 13-hydroxyglucopiericidin A.

8. The method for preparing 13-hydroxyglucopiericidin A according to claim 7, wherein: The purification comprises the following steps: The obtained fractions were purified by HPLC liquid chromatography, which included a 10 mm × 250 mm, 5 μm, YMC-pack ODS-A chromatographic column, a v / v mobile phase of acetonitrile:water = 45:55, and a flow rate of 2.5 mL / min.

9. The method for preparing 13-hydroxyglucopiericidin A according to claim 7, wherein: The Streptomyces psammoticus SCSIO NS126 is cultured on a solid state and expanded to obtain a fermentation product. The culture medium used for the solid culture comprises 6 g of yeast extract powder, 10 g of malt extract powder, 12 g of glucose, 20 g of agar, and 1 L of water. The pH is adjusted to 7.2 and the culture medium is sterilized. The culture medium for expanded culture is selected from a first culture medium or a second culture medium. The first culture medium includes 6 g of yeast extract powder, 10 g of malt extract powder, 12 g of glucose, 20 g of agar, and 1 L of water, and is adjusted to pH 7.2 and sterilized. The second culture medium includes 10 g of glucose, 3 g of yeast extract powder, 10 g of starch, 25 g of cottonseed powder, 2 g of NaCl, 5 g of CaCO3, and 1 L of water, and is adjusted to pH 4.0-7.2 and sterilized.

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