A sulfmyl-bridged phytocerberin compound, physalin S, and a preparation method and application thereof

Abstract

The present application provides a kind of containing sulfuryl bridge physalin compound small physalin S and its preparation method and application, from Physalis pubescens Franch. Isolated and extracted to the new compound small physalin S of physalin, it has the molecular structure of I:Small physalin S of the present application has anti-inflammatory activity, can significantly improve the viability of PC12 cell of hypoxia reperfusion injury, its mechanism of action is related to the expression of inhibiting NLRP3 inflammasome and cell pyroptosis related protein, inhibits the release of inflammatory factors, suggests that small physalin S of the present application can be used as the medicine popularization of treating ischemic brain injury, with good development, application prospect.

Description

Technical Field

[0001] This invention relates to the field of pharmaceutical technology, and in particular to a thionyl brines-containing physalin compound, small acid physalin S, its preparation method, and its application. Background Technology

[0002] Physalis alkekengi, belonging to the Solanaceae family and the Physalis genus, is an annual herb widely distributed and abundant in my country. The chemical composition of Physalis plants is complex, mainly including steroids, alkaloids, organic acids, and flavonoids, with physalin being a characteristic substance of this genus. Studies have isolated various chemical components such as physalin, phygrine, withaphysalin, and flavonoids from Physalis alkekengi. As a plant used for both food and medicine, Physalis alkekengi possesses strong physiological activity. Modern pharmacological studies have shown that physalin has good effects in antioxidation, anti-inflammation, hypoglycemia, antibacterial, analgesia, anticancer, uterine stimulation, labor induction, contraception, and diuresis. However, the extraction of physalin S from Physalis alkekengi, its preparation methods, and applications have not yet been reported. Summary of the Invention

[0003] In view of this, the present invention proposes a thionyl brines-containing physalin compound, small acid physalin S, its preparation method and application.

[0004] The technical solution of this invention is implemented as follows:

[0005] A compound containing thionyl brines, called small acid physalisin S, has the structure of formula I:

[0006]

[0007] A method for preparing a thionyl brittle bone compound, small acid bromelain S, comprising the following steps:

[0008] (1) Using the dried whole herb of Physalis alkekengi as raw material, add 9-11 times the mass of the raw material in 80-90 v / v% ethanol solution, reflux extract 2-4 times, each extraction for 1-3 hours, combine the extracts, concentrate under reduced pressure to recover the solvent, and obtain extract 1.

[0009] (2) Disperse extract 1 in 5-8 times its mass of water, filter and centrifuge, and precipitate the supernatant in a macroporous resin column using water-ethanol gradient elution with a volume ratio of 100:0-0:100. Recover the solvent under reduced pressure to obtain extract 2.

[0010] (3) The extract 2 from step (2) is suspended in water and extracted sequentially with petroleum ether, dichloromethane and ethyl acetate. The ethyl acetate extract is concentrated under reduced pressure to recover the solvent and obtain the ethyl acetate layer extract.

[0011] (4) The ethyl acetate extract was separated by silica gel column chromatography under reduced pressure, and eluted with a gradient of dichloromethane-methanol at a volume ratio of 20-1:1 to obtain 6 fractions: Fr.C-1, Fr.C-2, Fr.C-3, Fr.C-4, Fr.C-5 and Fr.C-6.

[0012] (5) Fraction Fr.C-3 was separated by vacuum MCI column silica gel column chromatography, and eluted with diethanol-water at volume ratios of 3:7, 6:4 and 9:1 to obtain fractions Fr.C-3-1, Fr.C-3-2 and Fr.C-3-3;

[0013] (6) The fraction Fr.C-3-2 was separated by Sephadex-LH column chromatography and isocratically eluted with methanol as the eluent to obtain fractions Fr.C-3-2A1, Fr.C-3-2A2 and Fr.C-3-2A3;

[0014] (7) The fraction Fr.C-3-2A1 was separated by semi-preparative high performance liquid chromatography and eluted isocratically with hexane-isopropanol in a volume ratio of 4:1 to obtain small acid syrup S as shown in Formula I.

[0015] Furthermore, in step (2), the filtration is performed using a 100-300 mesh filter cloth.

[0016] Furthermore, in step (2), the volume ratio of water to ethanol is 100:0, 70:30, 40:60, or 0:100.

[0017] Furthermore, in step (2), the elution portion of the vacuum recovery is a macroporous resin with a water-ethanol volume ratio of 40:60.

[0018] Furthermore, in step (4), the volume ratio of dichloromethane to methanol is 20:1, 15:1, 10:1, 5:1, 1:1, or 0:1.

[0019] Application of thionyl brittle acid-containing physalin compounds, specifically small acid physalin S, in the preparation of anti-inflammatory drugs.

[0020] Furthermore, the anti-inflammatory drug is an anti-inflammatory drug that reduces hypoxia-reperfusion injury in PC12 cells.

[0021] Compared with the prior art, the beneficial effects of the present invention are:

[0022] I. This invention isolates and extracts a new compound called small physalisin S from small physalis, whose molecular structure is a C(2)-C(6) six-membered ring system composed of thionyl groups in series.

[0023] II. The novel physalin compound, small physalin S, of the present invention has significant anti-inflammatory activity, reduces the release rate of lactate dehydrogenase and the release of NO, and alleviates hypoxia-reperfusion injury in PC12 cells.

[0024] III. The anti-inflammatory mechanism of the novel physalin compound small physalin S of the present invention is related to the inhibition of the expression of NOD-like receptor thermoprotein domain-associated protein 3, apoptosis-associated speckle-like protein, cleavage-type cysteine ​​protease-1, pyroptosis key protein, interleukin-18 and interleukin-1β. Attached Figure Description

[0025] Figure 1 The image shows the HRESIMS (mass spectrometry) spectrum of physalin S.

[0026] Figure 2 For small physalin S 1 H- 1 H COSY( 1 H- 1 H homonuclear correlation spectrum (C5D5N).

[0027] Figure 3 The HSQC (heteronuclear single quantum coherence spectrum) of small physalis S (C5D5N) is shown.

[0028] Figure 4 The HMBC (hydrogen detection heteronuclear multibond coherence spectrum) of small acid physalis S (C5D5N) is shown.

[0029] Figure 5 The ROESY (rotated coordinate NOE spectrum) of small physalin S (C5D5N).

[0030] Figure 6 The main component of small acid physalin S 1 H- 1 H COSY, HMBC, and ROESY related signals.

[0031] Figure 7 It is a single crystal structure of physalin S.

[0032] Figure 8 Cell viability was determined by administering different concentrations of physalin S (CCK-8 assay).

[0033] Figure 9 The effect of physalin S on LDH release rate in OGD / R-damaged PC12 cells.

[0034] Figure 10 The effect of physalin S on NO content in PC12 cells with OGD / R damage.

[0035] Figure 11The morphology of PC12 cells under an inverted microscope.

[0036] Figure 12 This study investigated the expression of NLRP3 (NOD-like receptor thermoprotein domain-associated protein 3), Cleaved-casp-1 (cleavage-type cysteine ​​protease-1), and ASC (apoptosis-associated speckle-like protein) in PC20 cells in vitro using syringin S.

[0037] Figure 13 This study investigated the expression of GSDMD-N (a key protein in pyroptosis), IL-18 (interleukin-18), and IL-1β (interleukin-1β) in PC20 cells in vitro using physalin S.

[0038] Figure 14 Fluorescent expression of GSDMD (a key protein of pyroptosis) in PC12 cells with OGD / R damage by physalin S. Detailed Implementation

[0039] To better understand the technical content of this invention, specific embodiments are provided below to further illustrate the invention.

[0040] Unless otherwise specified, the experimental methods used in the embodiments of this invention are all conventional methods.

[0041] Unless otherwise specified, all materials and reagents used in the embodiments of this invention are commercially available.

[0042] Example

[0043] (1) Using 20 kg of dried whole herb of Physalis alkekengi as raw material, add 10 times the mass of the raw material in 85% v / v% ethanol solution, reflux extract 3 times, extract for 2 hours each time, combine the extracts, concentrate under reduced pressure to recover the solvent, and obtain extract 1 (1.5 kg).

[0044] (2) Disperse extract 1 in 7 times its weight of water, filter it on a 200-mesh filter cloth, centrifuge, and chromatographically extract the supernatant into a macroporous resin column. Use water-ethanol with volume ratios of 100:0, 70:30, 40:60, and 0:100 as eluent gradients. Remove the solvent from the macroporous resin elution part with water-ethanol at a volume ratio of 40:60 under reduced pressure to obtain extract 2 (1 kg).

[0045] (3) The extract 2 from step (2) was suspended in water and extracted sequentially with petroleum ether, dichloromethane and ethyl acetate. The ethyl acetate extract was concentrated under reduced pressure to recover the solvent and obtain ethyl acetate layer extract (100g).

[0046] (4) The ethyl acetate extract was separated by silica gel column chromatography under reduced pressure, and eluted with dichloromethane-methanol gradient eluent at volume ratios of 20:1, 15:1, 10:1, 5:1, 1:1 and 0:1 to obtain 6 fractions: Fr.C-1, Fr.C-2, Fr.C-3 (13g), Fr.C-4, Fr.C-5 and Fr.C-6;

[0047] (5) Fraction Fr.C-3 (13g) was separated by vacuum MCI column silica gel column chromatography, and eluted with diethanol-water in volume ratios of 3:7, 6:4 and 9:1 to obtain fractions Fr.C-3-1, Fr.C-3-2 (2g) and Fr.C-3-3;

[0048] (6) The fraction Fr.C-3-2 (2g) was separated by Sephadex-LH column chromatography and isocratically eluted with methanol as the eluent to obtain fractions Fr.C-3-2A1 (300mg), Fr.C-3-2A2 and Fr.C-3-2A3;

[0049] (7) The fraction Fr.C-3-2A1 (300mg) was separated by semi-preparative high performance liquid chromatography. The chromatographic column was silica gel (5μm in diameter, 10×250mm in size). The elution was isocratic with hexane-isopropanol in a volume ratio of 4:1 to obtain small acid syrup S (16.80mg).

[0050] The small acid physalis S obtained in the examples 13 C NMR and 1 The H NMR data are shown in Table 1.

[0051] Table 1

[0052]

[0053] Note: 13 C NMR, 125MHz; 1 H NMR, 500MHz

[0054] Physicochemical properties of small acid phyll S: colorless needle crystals, mp 265-267℃ (dichloromethane:methanol = 1:1), (c 0.1, MeCN), 10% sulfuric acid ethanol solution appears orange-yellow on TLC, according to Figure 1 [MH] in the HRESIMS map - The quasi-molecular ion peak m / z is 591.1522 (calcd for 591.1536), and this is consistent with Table 1. 13 C10 NMR data determined the molecular formula of physalin S to be C10. 28 H 32 O 12S.

[0055] As can be seen from Table 1, 13 δ in the low-field region of C NMR C 200.7(C-1) and δ C 208.3 (C-15) represents the signals of the two ketone carbonyl carbons, δ C 173.4 (C-18) and δ C 167.4 (C-26) represents the two carbonyl carbons of the ester, δ C 107.1 (C-14) is a typical ketal signal. Based on the physicochemical properties of the compound, it is inferred that small physalin S may be a type IA physalin compound.

[0056] according to Figure 2 of 1 H- 1 The H-COSY spectrum shows correlation signals between H-2 / H-3 and H-3 / H-4a, and also combines... Figure 3 The HSQC signal indicates that C-2 / C-3 / C-4 are a group of adjacent carbon signals. From Figure 4 The HMBC spectrum clearly shows a strong correlation between the H-3 and C-1 ketone carbonyl groups, while the H-2 and C-1 ketone carbonyl groups show a weaker correlation. Based on the structural characteristics of the A ring of physalin compounds, the possible structure of the A ring of physalin S can be roughly attributed. Figure 4 The HMBC spectrum also shows the correlation between Me-19 and C-5. Combined with the relatively low field chemical shift at C-5, it suggests that C-5 is a quaternary carbon with a hydroxyl substitution, similar to common physalin compounds.

[0057] according to Figure 4 The HMBC spectrum showed weak correlation signals between H-6 and C-2, and between H-2 and C-6, as well as correlation signals between H-2, H-6, and C-4, suggesting the possible existence of a rare C(2)-C(6) cyclic system in this compound. This, combined with the chemical shifts at C-2 and C-6 and the specific chemical shift (δ) of the ketone carbonyl group at C-1, further supports this finding. C-1 (200.7 ppm), if there are no other conjugated systems in ring A, the chemical shift of the ketone carbonyl group at position C-1 should be greater than 210.0 ppm, combined with the molecular formula C 28 H 32 O 12Based on information such as S and unsaturation, a special thionyl ring structure with a C(2)-C(6) bridge formed by sulfur atoms (S) in series may have been formed here, creating a 6-membered ring system with the molecule's parent nucleus. Simultaneously, the large γ-effect of the thionyl group causes the chemical shift at C-1 to shift significantly to a higher field. This reasonably explains the special chemical shift of the C-1 ketone carbonyl group, because if there is no double-bonded conjugated system in ring A, the chemical shift of the C-1 ketone carbonyl group would be abnormally low, around 10.0 ppm.

[0058] The chemical shift at position C-19 in physalin S is δ C 8.0 ppm (<10.0 ppm) suggests that rings A and B are cis-coupled, meaning the 5-OH group is in the β configuration. From Figure 5 The ROESY spectrum showed correlation signals between H-2 and H-6 and H-4β, suggesting that the anomalous six-membered ring is of α configuration. Figure 6 The main component of small acid physalin S 1 H- 1 H COSY, HMBC, and ROESY related signals, at the same time Figure 7 The single-crystal data of this compound proves that it is the first isolated physalis compound with a C(2)-C(6) six-membered ring system containing thionyl groups in series. It also gives the stereoconfiguration of physalis S as 2S, 3R, 5S, 6S, 8R, 9S, 10R, 13R, 14R, 16S, 17R, 20S, 22R, 24S, 25S.

[0059] In summary, based on relevant data from the literature, this compound is a new compound, named Physalis minor S, and its structure is shown in Formula I:

[0060]

[0061] Anti-inflammatory activity test

[0062] 1. Experimental groups: blank control group, oxygen-glucose deprivation / reoxygenation (OGD / R) group, and drug administration group (OGD / R + small acid syrup S).

[0063] 2. Modeling and Drug Administration: Rat adrenal pheochromocytoma cells (PC12) were purchased from the Shanghai Cell Bank of the Chinese Academy of Sciences. PC12 cells in the logarithmic growth phase were used at a dose of 1×10⁻⁶. 5 Cells were seeded at a density of 1 cell / mL in 96-well plates.

[0064] Blank control group: PC12 cells were kept in DMEM medium containing 10% fetal bovine serum and 1% penicillin and cultured in a cell culture incubator at 37°C and 5% CO2.

[0065] Oxygen-glucose deprivation / reoxygenation (OGD / R) group: PC12 cells seeded in 96-well plates were cultured for 32 h. The cell culture medium of the OGD / R group was replaced with glucose-free Earle's balanced salt solution and placed in a three-gas incubator with a volume fraction of 95% N2-4% CO2-1% O2 for 3 h of glucose- and oxygen-deficient culture. Then the cell culture medium was replaced with DMEM medium containing 10% fetal bovine serum and 1% penicillin antibiotics and placed in a cell culture incubator at 37°C and 5% CO2 for reoxygenation for 12 h.

[0066] The treatment group (OGD / R + small acid syrup S): After culturing PC12 cells in 96-well plates for 24 hours, a certain concentration of small acid syrup S solution was added to the treatment group. The small acid syrup S solution was prepared using serum-free DMEM medium. After culturing for another 8 hours, the cell culture medium in the treatment group was replaced with glucose-free Earle's balanced salt solution, and the cells were placed in a three-gas incubator with a volume fraction of 95% N2-4% CO2-1% O2 for 3 hours of glucose- and hypoxia-deficient culture. Subsequently, the cell culture medium was replaced with DMEM medium containing 10% fetal bovine serum and 1% penicillin antibiotics, and the cells were placed in a cell culture incubator at 37°C and 5% CO2 for reoxygenation and perfusion for 12 hours.

[0067] 3. Determination of the optimal concentration of small acid lysin S: Cell viability assay (CCK-8 assay): PC12 cells in logarithmic growth phase were inoculated at 1×10⁻⁶ cells / year. 5 Cells were seeded at a concentration of 100 μL / mL into 96-well plates, with 5 replicates per group. After 24 h of culture, the treatment groups were treated with 2.5 μM, 5 μM, 10 μM, 20 μM, 40 μM, 80 μM, and 160 μM of small acid lysine S solution, respectively, and incubated at 37°C in a 5% CO2 cell culture incubator for another 12 h. The 96-well plates were then removed, and 10 μL of CCK-8 reagent was added to each well under dark conditions. The plates were incubated for another 30 min, and the absorbance (A) was measured at 450 nm. The viability of PC12 cells was calculated using the following formula:

[0068] Cell viability = (A experimental wells - A blank wells) / (A blank control wells - A blank wells)

[0069] The experimental wells include the OGD / R group and the drug administration groups with different concentrations of physalin S solution. The blank wells represent cell-free culture medium wells, and the blank control wells represent wells that have not been treated with OGD / R modeling or physalin S solution.

[0070] CCK-8 test results are as follows Figure 8As shown, compared with the blank control group, the PC12 cell viability in the OGD / R group was significantly decreased (P<0.01). Compared with the OGD / R group, different concentrations of physalin S significantly improved PC12 cell viability (P<0.01, P<0.05), with the 80 μM concentration showing the most significant improvement. Therefore, for subsequent PC12 cell anti-inflammatory activity tests, 20 μM, 40 μM, and 80 μM were selected as the optimal concentrations for physalin S administration, with 80 μM being the optimal concentration.

[0071] 4. Detection indicators

[0072] Lactate dehydrogenase (LDH) is a stable cytoplasmic enzyme present in most cells. When cells are damaged, it can be released into the culture medium. Therefore, the role of cytoplasmic acid S can be assessed by detecting the LDH release rate in OGD / R-damaged PC12 cells using a cytotoxicity assay kit. The luciferin substrate Cytotox 96 assay kit is added to the cell culture medium. When the cell membrane is damaged, LDH is released into the medium, reacting with the substrate to generate a fluorescent signal. The fluorescence intensity is read using a fluorescence plate reader, and the LDH release rate is calculated. The detection results are as follows: Figure 9 As shown, from Figure 9 It can be seen that, compared with the blank control group, the LDH release rate of PC12 cells in the OGD / R group was significantly increased (P<0.01), and compared with the OGD / R group, the LDH release rate of the drug-treated groups (P<0.05) was significantly decreased (P<0.05). The results indicate that Phenolic acid S can alleviate OGD / R damage to PC12 cells.

[0073] Nitric oxide (NO) is an important messenger and effector molecule in the body. It is released when cells are damaged, exhibiting neurotoxicity, and is also a regulator of inflammatory responses. Therefore, the role of physalin S was assessed by detecting the NO content in the supernatant of OGD / R-damaged PC12 cell culture medium using the Griess method. The results are as follows: Figure 10 As shown, from Figure 10 It can be seen that, compared with the blank control group, the NO content of PC12 cells in the OGD / R group was significantly increased (P<0.01), and compared with the OGD / R group, the NO content of the drug-treated groups (P<0.01) was significantly decreased (P<0.01). The results indicate that P-P-S can reduce the release of the inflammatory factor NO and alleviate OGD / R damage to PC12 cells.

[0074] Figure 11 Images of PC12 cell morphology under an inverted microscope, from Figure 11It can be seen that the PC12 cells in the blank control group are polygonal in shape with long synapses, exhibiting dendritic characteristics of nerve cells. Compared with the blank control group, the PC12 cells in the OGD / R group are shrunken, rounded in shape, and have fewer adherent cells. Compared with the OGD / R group, in the drug treatment groups (Physalisin S concentrations of 20 μM, 40 μM, and 80 μM), the PC12 cell morphology gradually tends to normal spindle shape with an increase in adherent cells as the drug concentration increases. The results indicate that Physalisin S can significantly alleviate OGD / R damage to PC12 cells.

[0075] 5. Mechanism of action

[0076] Experimental Methods: Protein expression was detected by Western blot: 1. Protein extraction: 100 mg of brain tissue and 1 mL of RIPA tissue lysis buffer containing protease inhibitors and phosphatase inhibitors were added to a pre-chilled glass homogenizer. The cells were homogenized on ice for 20 min to fully lyse them. The cells were centrifuged for 15 min (12000 rpm, 4℃), and the supernatant was collected and placed in an EP tube for later use; 2. Total protein concentration was detected by BCA method: 1 μL of protein supernatant and 19 μL of deionized water were added to each well of a 96-well plate to prepare standard curves. BCA was then added to the standard curves. 1. **Incubation at 37℃ for 30 min:** Measure absorbance at 562 nm. 2. **Boiled Protein:** Add 1 / 4 volume of SDS buffer to the EP tube containing the protein supernatant and mix well. Cover the EP tube with sealing film and boil in 100℃ water for 10 min. Seal and store at -80℃ for later use. 3. **Separating Gel:** Prepare the optimal concentration of separating gel based on the molecular weight of the target protein. Gently mix using a pipette, then transfer the mixture to the gap between glass plates. Add pure water for water sealing. After the separating gel solidifies, add the prepared stacking gel and immediately insert it into the comb well. Note that no air bubbles should be generated throughout the process; 5. Electrophoresis: Use a pipette to transfer the pre-calculated volume of protein sample into the sample loading tank, fix it in the electrophoresis tank, add an appropriate amount of electrophoresis working solution to the electrophoresis tank, turn on the electrophoresis apparatus, run at 75V for about 30 minutes until the marker is fully broken in, then switch to 115V constant voltage electrophoresis for 60 minutes; 6. Transfer: Activate the PVDF membrane in methanol, remove excess gel, transfer it to a sandwich-like transfer plate, place the PVDF membrane (without air bubbles) on top, embed it in the transfer tank, and add 200 ml of transfer buffer. 7. Transferring protein onto a PVDF membrane using constant current (mA) for 2 hours; 8. Blocking: After transfer, wash the membrane three times with TBST for 5 minutes each time, then add rapid blocking buffer for blocking; 9. Primary antibody incubation: After blocking, wash the PVDF membrane three times with TBST for 5 minutes each time. After washing, place the PVDF membrane in an antibody incubation box containing the corresponding primary antibody, label it, and incubate overnight at 4°C on a shaker. The next day, recover the primary antibody, wash the PVDF membrane three times with TBST, and then incubate it for 2 hours in an antibody incubation box containing the corresponding species secondary antibody. The corresponding antibody concentrations are shown in Table 2. 10. Exposure: After antibody incubation, wash the PVDF membrane three times with TBST. During this time, prepare the ECL chemiluminescence solution (solution A:solution B ratio 1:1). After washing, place the membrane face up in the dark chamber of the gel imaging system, add the chromogenic solution, expose, and save the data. Analyze the experimental results using ImageJ software. The experimental results are shown in Table 2. Figure 12 , 13 As shown.

[0077] Table 2

[0078]

[0079] from Figure 12 As can be seen, compared with the blank control group (NC), the expression levels of NLRP3, Cleaved-casp-1, and ASC proteins in PC12 cells of the OGD / R group were significantly increased (P<0.001). Compared with the OGD / R group, the drug-treated groups (Plasmin S concentrations of 20 μM, 40 μM, and 80 μM) significantly reduced the expression levels of NLRP3, Cleaved-casp-1, and ASC proteins (P<0.05, P<0.01). Figure 13 As can be seen, compared with the blank control group (NC), the expression of GSDMD-N, IL-18, and IL-1β proteins in PC12 cells of the OGD / R group was significantly increased (P<0.01). Compared with the OGD / R group, the drug-treated groups (20 μM, 40 μM, and 80 μM of chlorpromazine S) significantly reduced the expression of GSDMD-N, IL-18, and IL-1β proteins (P<0.01). The results indicate that chlorpromazine S can inhibit the expression of NLRP3, GSDMD-N, IL-18, and IL-1β.

[0080] Figure 14 The effect of phylloidin S on the fluorescence expression of GSDMD protein in OGD / R-damaged PC12 cells was detected using cellular immunofluorescence. The figures show that, compared with the blank control group, the fluorescence expression of GSDMD protein in PC12 cells of the OGD / R group was significantly increased. Compared with the OGD / R group, the drug-treated group (phylloidin S concentration 80 μM) significantly reduced the fluorescence intensity of GSDMD protein in PC12 cells. These results indicate that phylloidin S has an inhibitory effect on the expression of GSDMD, a key protein in pyroptosis.

[0081] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of the present invention should be included within the protection scope of the present invention.

Claims

1. A thionyl brittle bone compound containing small acid physalis S, characterized in that, It has the structure of Formula I: I 。 2. The extraction method for a thionyl brittle bone compound containing small acid physalis S as described in claim 1, characterized in that, Includes the following steps: (1) Using the dried whole herb of Physalis alkekengi as raw material, add 9-11 times the mass of the raw material in 80-90 v / v% ethanol solution, reflux extract 2-4 times, each extraction for 1-3 hours, combine the extracts, concentrate under reduced pressure to recover the solvent, and obtain extract 1. (2) Disperse extract 1 in 5-8 times its mass of water, filter and centrifuge, and perform macroporous resin column chromatography on the supernatant. Use water-ethanol with volume ratios of 100:0, 70:30, 40:60 and 0:100 as eluent for gradient elution. Remove the solvent from the macroporous resin elution part with water-ethanol at a volume ratio of 40:60 to obtain extract 2. (3) The extract 2 from step (2) is suspended in water and extracted sequentially with petroleum ether, dichloromethane and ethyl acetate. The ethyl acetate extract is concentrated under reduced pressure to recover the solvent and obtain the ethyl acetate layer extract. (4) The ethyl acetate extract was separated by silica gel column chromatography under reduced pressure, and eluted with a gradient of dichloromethane-methanol at a volume ratio of 20-0:1 to obtain 6 fractions: Fr.C-1, Fr.C-2, Fr.C-3, Fr.C-4, Fr.C-5 and Fr.C-6. (5) Fraction Fr.C-3 was separated by vacuum MCI column silica gel column chromatography, and eluted with diethanol-water in volume ratios of 3:7, 6:4 and 9:1 to obtain fractions Fr.C-3-1, Fr.C-3-2 and Fr.C-3-3; (6) The fraction Fr.C-3-2 was separated by Sephadex-LH column chromatography and eluted isocratically with methanol as the eluent to obtain fractions Fr.C-3-2A1, Fr.C-3-2A2 and Fr.C-3-2A3; (7) The fraction Fr.C-3-2A1 was separated by semi-preparative high performance liquid chromatography and eluted isocratically with hexane-isopropanol in a volume ratio of 4:1 to obtain small acid syrup S as shown in Formula I.

3. The method for preparing a thionyl brittle bone compound containing small acid physalis S as described in claim 2, characterized in that, In step (2), the filtration is performed using a 100-300 mesh filter cloth.

4. The method for preparing a thionyl brittle bone compound containing small acid brittle bone syrup S as described in claim 2, characterized in that, In step (4), the volume ratio of dichloromethane to methanol is 20:1, 15:1, 10:1, 5:1, 1:1, or 0:

1.

5. The use of the thionyl brines-containing physalin compound, small acid physalin S, as described in claim 1, in the preparation of an anti-inflammatory drug to alleviate hypoxia-reperfusion injury in PC12 cells.

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