CN115040502B - Application of marine aspergillus source compound - Google Patents
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Abstract
The invention belongs to the technical field of biological medicines, and particularly relates to application of a marine aspergillus source compound in preparation of a medicine for preventing and treating non-alcoholic fatty liver disease. Cell experiments and animal experiments prove that the marine aspergillus source compound has obvious effect of treating the non-alcoholic fatty liver, and provides more choices for treating the non-alcoholic fatty liver; the compound is produced by utilizing the marine aspergillus with rich sources and combining with the modern microbial fermentation engineering technology, has the advantages of no worry of raw materials after the production, no damage to ecological balance, easy realization of industrialization and the like, is a renewable medicine resource with development prospect, and provides rich raw materials for researching and developing marine microbial products and marine microbial medicines.
Description
Technical Field
The invention belongs to the technical field of biological medicine, and relates to a new application of a known compound. More particularly relates to application of a natural marine aspergillus source compound in preparing a medicine for preventing and treating non-alcoholic fatty liver disease.
Background
Nonalcoholic fatty liver (NAFLD) is a metabolic liver disease characterized mainly by excessive accumulation of liver lipid (steatosis), and can progress gradually to nonalcoholic steatohepatitis (NASH), liver fibrosis, cirrhosis and hepatocellular carcinoma. NASH is considered as a key determining link in the progress of chronic liver disease, and is an irreversible disease between cirrhosis and liver cancer precursor lesions. Currently, the number of worldwide NAFLD/NASH patients is dramatically increased, and the number of NASH patients exceeds that of hepatitis B as the first major type of chronic liver disease worldwide, which is a global major public health problem.
In the prior art, NAFLD/NASH treatment methods are very limited, diet control and exercise intervention are mainly used for NAFLD and early NASH treatment, and the treatment effects of operations such as clinical weight loss and liver transplantation are general for middle and late NASH. In the aspect of drug therapy, researchers find a plurality of therapeutic targets and targeted drugs thereof aiming at the molecular activities of various important cells of NASH, and clinically developed NASH therapeutic drugs are mainly divided into: farnesol receptor (FXR) agonists, peroxisome proliferator-activated receptor (PPAR) agonists, acetyl-coa carboxylase (ACC) inhibitors, apoptosis enzyme inhibitors, and the like. For example, chinese patent application discloses a compound for modulating FXR, which has a good effect of modulating farnesol receptor. In addition, as FXR agonist-like drug obeticholic acid, it is used for treating primary biliary cirrhosis and nonalcoholic fatty liver disease by indirectly inhibiting cytochrome 7A1 (CYP 7A 1) gene expression, thereby inhibiting cholic acid synthesis; however, problems of efficacy (not effective against all critical cellular molecular activities of NASH), safety (itching, weight gain, heart failure, etc.) have been successively terminated in clinical trials. In addition, the safety and long-term efficacy of the compounds under clinical investigation remain to be further verified.
Therefore, in order to solve the shortage problem of the existing NASH therapeutic drugs, the research and development of the drugs with high efficiency, low toxicity, high safety and good NAFLD/NASH resisting effect is significant.
Disclosure of Invention
The invention aims to overcome the defects and shortcomings of the existing NAFLD therapeutic drug with high safety and obvious effect, and provides the application of the natural marine aspergillus source compound with high safety and good NAFLD therapeutic effect in preparing the drug for preventing and treating the non-alcoholic fatty liver disease.
It is another object of the present invention to provide a process for the preparation of said marine aspergillus source compound.
The above object of the present invention is achieved by the following technical scheme:
the application of a marine aspergillus source compound in preparing a medicament for preventing and treating non-alcoholic fatty liver disease is characterized in that the marine aspergillus source compound has a structure of a formula (I):
further, the non-alcoholic fatty liver includes inhibiting liver lipid accumulation, liver injury, liver inflammation and fibrosis.
Still further, the marine aspergillus source compound also includes pharmaceutically acceptable salts, hydrates, solvates thereof.
Further, the medicament for preventing and treating the nonalcoholic fatty liver disease is in the form of an oral preparation, an injection or an inhalant.
In addition, the invention also provides a preparation method of the marine aspergillus source compound, wherein the marine aspergillus source compound is obtained by fermenting and separating marine aspergillus;
the marine aspergillus source compound has a structure of formula (I):
further, the marine aspergillus is a coanda fungus separated from marine hard coral.
Still further, the marine Aspergillus comprises Aspergillus sp.ITBB-c1, aspergillus taichungensis ZHN-7-07, aspergillus candidus.
Further, the preparation method specifically comprises the following steps:
s1, performing expansion culture on marine aspergillus, inoculating strain liquid into a culture medium containing rice, and fermenting to obtain fermentation liquor;
s2, leaching the fermentation liquor obtained in the step S1 by using ethyl acetate, concentrating the leaching liquor, and sequentially carrying out pressure-reducing normal-phase silica gel column chromatographic separation by using a petroleum ether-ethyl acetate system as an eluent according to the volume ratio of petroleum ether to ethyl acetate of 100:0, 25:1, 10:1, 5:1, 2:1, 1:1 and 0:1 to obtain 7 fractions from Fr.A to Fr.G;
s3, collecting fraction Fr.E obtained in the step S2 with the volume ratio of petroleum ether to ethyl acetate being 2:1, and carrying out ODS reversed-phase column chromatography by taking 40-100 vol% methanol water solution as an eluent;
s4, collecting an eluted part of the 75vol% methanol aqueous solution in the step S3 to obtain a fraction Fr.E-6, using methanol as an eluent, completely carrying out Sephadex LH-20 gel column chromatography, and concentrating to obtain the marine aspergillus source compound.
Further, in the step S1, the fermentation condition is that the fermentation is carried out at 25-30 ℃ for 40-50 days.
Further, in the step S1, the culture medium containing rice is prepared from cooked rice and water according to the mass volume ratio of 1 (1-1.5) g/ml.
The invention has the following beneficial effects:
the invention provides an application of a marine aspergillus source compound in preparing a medicament for preventing and treating non-alcoholic fatty liver, and cell experiments and animal experiments prove that the marine aspergillus source compound has obvious effect of treating the non-alcoholic fatty liver and provides more choices for treating the non-alcoholic fatty liver; the compound is produced by utilizing the marine aspergillus with rich sources and combining with the modern microbial fermentation engineering technology, has the advantages of no worry of raw materials after the production, no damage to ecological balance, easy realization of industrialization and the like, is a renewable medicine resource with development prospect, and provides rich raw materials for researching and developing marine microbial products and marine microbial medicines.
Drawings
FIG. 1 is a graph showing the hydrogen spectrum of the marine koji mold-derived compound HN-001 obtained in example 1 of the present invention 1 H-NMR，500MHz)。
FIG. 2 is a mass spectrum of the marine koji mold source compound HN-001 obtained in example 1 of the present invention.
FIG. 3 is a photograph of Nile red stained cells (FIG. 3B) at the level of intracellular Triglyceride (TG) (FIG. 3A) and representative concentrations (4-25. Mu.M) in example 2 of the present invention.
FIG. 4 is a graph showing the therapeutic effect of the compound of example 3 of the present invention on palmitic acid-induced hepatocyte apoptosis (FIG. 4A) and the morphology change of each group of hepatocytes (FIG. 4B) and the activity level of each group of apoptotic enzymes (FIG. 4C).
FIG. 5 is a statistical plot of the effect of the compounds of example 4 of the present invention on the active oxygen content of palmitic acid-induced hepatocyte lipid oxidation stress markers.
FIG. 6 is a graph of the weight change curve of the mouse model in example 5 of the present invention (FIG. 6A), a statistical graph of the weight data of the mouse after 7 weeks of administration (FIG. 6B), and a liver/weight comparison graph of the mouse model after 7 weeks of administration (FIG. 6C).
FIG. 7 is a statistical chart of the content levels of triglyceride (FIG. 7A) and glutamic-oxaloacetic transaminase (AST), glutamic-pyruvic transaminase (ALT) and alkaline phosphatase (ALP) (FIG. 7B) in the mouse model liver tissue according to example 5 of the present invention; and liver histopathological staining patterns of the mouse model, including hematoxylin/eosin staining, oil red O staining, sirius red staining, and masson staining (fig. 7C).
FIG. 8 shows the result of cytotoxicity test of HN-001 against hepatocytes of the compound of example 6 of the present invention.
Detailed Description
The invention is further illustrated in the following drawings and specific examples, which are not intended to limit the invention in any way. Unless specifically stated otherwise, the reagents, methods and apparatus employed in the present invention are those conventional in the art.
The marine Aspergillus sp.ITBB-c1 adopted in the invention is obtained by separating marine hard coral.
Reagents and materials used in the following examples are commercially available unless otherwise specified.
Example 1 preparation of Marine Aspergillus source Compound HN-001
The preparation of the marine aspergillus source compound HN-001 comprises the following steps:
s1, activating a marine Aspergillus sp.ITBB-c1 strain by a PDA solid culture medium plate, and performing dark culture for 5 days at a culture condition of 28 ℃; directly inoculating colony blocks in a flat plate into a 1000mL triangular flask filled with 200mL ME seed culture medium, and performing shake culture on a constant temperature shaking table for 4 days (160 r/min,28 ℃) to obtain seed liquid; inoculating 15mL of seed solution into a tissue culture bottle filled with 30g of rice and 30mL of water for fermentation, fermenting 5kg of rice in total, and standing and culturing at 28 ℃ for 45 days to obtain fermentation liquor;
s2, leaching the fermentation liquor obtained in the step S1 with ethyl acetate for 4 times, combining the leaching liquor and concentrating under reduced pressure at 45 ℃ to obtain 120g of ethyl acetate extract; adopting a petroleum ether-ethyl acetate system as an eluent, sequentially carrying out reduced pressure normal phase silica gel column chromatographic separation according to the volume ratio of petroleum ether-ethyl acetate of 100:0, 25:1, 10:1, 5:1, 2:1, 1:1 and 0:1 (gradient elution), and combining similar fractions under TLC detection to obtain 7 fractions from Fr.A to Fr.G;
s3, collecting fraction Fr.E obtained in the step S2 with the volume ratio of petroleum ether to ethyl acetate being 2:1, and carrying out ODS reversed-phase column chromatography by taking 40-100 vol% methanol water solution as an eluent;
s4, collecting an eluted part of the 75vol% methanol aqueous solution in the step S3 to obtain a fraction Fr.E-6, using methanol as an eluent, completely carrying out Sephadex LH-20 gel column chromatography, and concentrating to obtain the marine aspergillus source compound HN-001 (2.0 mg).
FIGS. 1 to 2 are structure confirmation maps of marine Aspergillus source compounds HN-001: it can be seen that positive source high resolution mass spectrometry (HR-ESI-MS) excimer ion peak of compound HN-001: m/z 381.1336[ M+H ]]+，783.2416[2M+Na]The push-out molecular formula is C 22 H 20 O 6 。
Example 2 influence of Marine aspergillus source compound HN-001 on palmitic-induced hepatocyte lipid content (nile red staining)
In the embodiment, a compound HN-001 derived from marine aspergillus is taken as a test object, and the influence of HN-001 on the triglyceride content in liver cells is verified by adopting a nile red staining method.
1. The experimental method comprises the following steps:
HepG2 cells in logarithmic growth phase were taken according to 5.0X10 4 Cells/well, plated evenly into 48-well plates, and cultured overnight. The medium was changed to a complete DMEM medium (10% FBS and 1% diabody) containing palmitic acid (0.5 mM), and various concentrations (4, 10, 25. Mu.M) of compound HN-001 diluted with complete DMEM medium containing palmitic acid, 37℃and 5% CO were added 2 And (5) standing and culturing for 24 hours. The experiments set up a blank control group, a palmitic acid model group and a compound HN-001 treatment group. After the end of 24 hours of action, the groups were photographed on nile red staining and analyzed for triglyceride content.
The specific operation of the nile red dyeing is as follows: the cells to be stained were rinsed 1 time with pre-chilled PBS, added with 1. Mu.M nile red staining solution and 10. Mu.M Hochest 33342 nucleic acid dye, and stained at room temperature for 15 minutes. After dyeing is completed, rinsing with deionized water for 2-3 times at room temperature, and photographing with an inverted microscope (40 times).
The triglyceride content analysis experiment comprises the following specific steps: the cells to be detected were rinsed 2 times with pre-chilled PBS. Removing PBS, adding deionized solution containing 0.2% Triton X-100, standing at room temperature for 1 hr, collecting cell suspension, ultrasonic crushing for 10min to allow cells to be fully lysed, centrifuging, collecting supernatant, and determining triglyceride content according to triglyceride detection kit (Roche) instruction.
Wherein, the triglyceride content analysis uses palmitic acid control group as control (Ctrl), and the fold relationship is:
the experimental results are the average of three independent experiments, and the results are statistically analyzed according to the average value + -standard deviation.
2. Experimental results:
as a result of the experiment, referring to FIG. 3, it can be seen that the compound HN-001 can reduce the triglyceride content of the liver cells in a gradient-dependent manner at the concentration of 4-25 mu M. The results of the triglyceride quantitative assay showed that the use of compound HN-001 can reduce intrahepatic triglyceride levels (fig. 3A). Nile red staining further demonstrated the lipid lowering activity of compound HN-001: the large amount of lipid was clearly visible in the cells of the control group, while the lipid content was significantly reduced in the cells of the compound HN-001 treated group (fig. 3B).
Example 3 protection of Marine aspergillus source compound HN-001 against palmitic acid-induced apoptosis of hepatocytes (trypan blue staining)
In the embodiment, a compound HN-001 derived from marine aspergillus is taken as a test object, and a trypan blue staining method is adopted to verify the study on the protective effect of the compound HN-001 on palmitic acid induced hepatocyte apoptosis.
1. The experimental method comprises the following steps:
HepG2 cells in logarithmic growth phase were taken at 20X 10 4 Cells/well, plated evenly in 6-well plates, and cultured overnight. The medium was changed to a complete DMEM medium (10% FBS and 1% diabody) containing palmitic acid (0.5 mM), and various concentrations (4, 10, 25. Mu.M) of compound HN-001 diluted with complete DMEM medium containing palmitic acid, 37℃and 5% CO were added 2 And (5) standing and culturing for 24 hours. The experiments set up a blank control group, a palmitic acid model group and a compound HN-001 treatment group. After 24 hours of completion of the action, cell survival and apoptosis were detected by trypan blue staining and Caspase3 (Caspase 3) activity was detected in each group of cells using the kit.
The specific operations of trypan blue staining are: cells to be stained were rinsed 1 time with pre-chilled PBS, collected cells by pancreatin digestion, resuspended in 1mL of phosphate buffer solution containing 4% trypan blue staining solution, and stained for 15 minutes at room temperature. After staining was completed, 10 μl of the cell suspension was taken into a cell counting plate, and the cell staining was counted under a microscope, and the numbers of trypan blue stained and undyed cells of each group were recorded, respectively. The experimental results are the average of three independent experiments, and the results are statistically analyzed according to the average value + -standard deviation.
The specific operation of the Caspase3 activity detection is as follows: after the cell treatment was completed, the cells were lysed on chilled cell lysates on ice for 15 minutes. 10 microliters of sample supernatant was added to the Caspase3 antibody coated microplate and incubated at 37℃for 1 hour. Unbound specimens were discarded and the plates were washed 3 times for 1 minute each with incubation buffer at room temperature. Adding a freshly prepared Caspase substrate solution, and reacting for 3 hours at 37 ℃; then fluorescence photometry is carried out, the excitation wavelength is 400nm, and the emission wavelength is 505nm.
2. Experimental results:
as a result, referring to fig. 4, it can be seen from the graph that compound HN-001 can gradient-dependently increase hepatocyte survival at a concentration of 4 to 25 μm, and decrease palmitic acid-induced apoptosis of hepatocytes (fig. 4A). Cell morphology staining further demonstrated the hepatocyte protective activity of compound HN-001. Compound HN-001 treated group had more regular intracellular morphology and reduced numbers of shrunken cells compared to the palmitic acid group cells (fig. 4B). At the same time, palmitic acid induction significantly increased hepatocyte Caspase3 activity, whereas compound HN-001 treatment significantly reduced palmitic acid-induced hepatocyte Caspase3 activity (fig. 4C).
Example 4 inhibition of palmitic acid-induced oxidative stress of hepatocytes by Marine aspergillus source compound HN-001 (flow cytometry to detect cellular Reactive Oxygen Species (ROS) content)
In this example, the influence of compound HN-001 on the level of palmitic acid-induced hepatocyte active oxygen was examined by flow cytometry using compound HN-001 derived from marine Aspergillus as a test subject.
3. The experimental method comprises the following steps:
HepG2 cells in logarithmic growth phase were taken at 20X 10 4 Cells/well, plated evenly in 6-well plates, and cultured overnight. The medium was changed to a complete DMEM medium (10% FBS and 1% diabody) containing palmitic acid (0.5 mM), and various concentrations (4, 10, 25. Mu.M) of compound HN-001 diluted with complete DMEM medium containing palmitic acid, 37℃and 5% CO were added 2 And (5) standing and culturing for 24 hours. The experiments set up a blank control group, a palmitic acid model group and a compound HN-001 treatment group. After 24 hours of completion of the action, the cells were collected and assayed for cellular reactive oxygen species by flow cytometry.
The active oxygen level detection specifically operates as: cells to be stained were rinsed 1 time with pre-chilled PBS, collected cells by pancreatin digestion, resuspended in 1mL of medium, and stained with 10. Mu.M DCFH-DA active oxygen probe at 37℃for 15 minutes. After staining, cells were collected by centrifugation, resuspended in 500. Mu.L of phosphate buffer, and the intracellular levels of active oxygen were measured by flow cytometry. The experimental results are the average of three independent experiments, and the results are statistically analyzed according to the average value + -standard deviation.
4. Experimental results:
as a result, referring to fig. 5, it can be seen from the graph that Palmitic Acid (PA) induction can significantly increase the level of active oxygen in hepatocytes, and that compound HN-001 can decrease the level of active oxygen in hepatocytes in a gradient-dependent manner at a concentration of 4 to 25 μm (fig. 5).
Example 5 Effect of Marine Aspergillus source Compound HN-001 on non-alcoholic fatty liver
In this example, in vivo validation experiments were performed on a high-fat high-cholesterol diet-induced non-alcoholic fatty liver mouse model with a marine Aspergillus sp.c1-derived compound HN-001 test subject.
1. The experimental method comprises the following steps:
(1) Constructing a non-alcoholic fatty liver model:
male C57BL/6J mice (18-20 g in body weight) of 8 weeks old were fed high fat high cholesterol Diet (HFC, 60% fat and 1.2% cholesterol, available from Research Diet Inc. of America) or normal Diet (available from laboratory animal center of Guangdong) for 10 weeks.
(2) Preparing a compound HN-001 solution:
a physiological saline solution containing 2.5% PEG-400 and 5% castor oil is used for preparing a compound HN-001 solution with the concentration of 2mg/mL, and the compound HN-001 solution is fully mixed and dissolved.
(3) Vehicle or compound HN-001 intervention:
HFC dietary mice were randomly divided into two groups (HFC model control group and HN-001 dosing group), 10 each. Normal feed mice continue to be fed with normal feed, HFC model control group and HN-001 group mice continue to be fed with HFC feed. Wherein, HN-001 group is given the HN-001 solution prepared in the step (2) by intraperitoneal injection according to the dosage of 20mg/kg, and the control group is given the solvent treatment. Once every two days, mice were observed and recorded for food intake and body weight. All mice were anesthetized, bled 7 weeks after dosing, and cervical dislocation sacrificed and dissected and each group of mice weight and liver weight recorded.
(4) Analysis of mouse blood samples:
centrifuging the mouse blood sample collected in the step (3) at 3000 rpm for 10min, and taking a supernatant. Detecting indexes such as glutamic-oxaloacetic transaminase, glutamic-pyruvic transaminase and the like in serum.
(5) Mouse liver tissue sample analysis:
weighing a certain mass of the liver tissue of the mice collected in the step (3), crushing the tissue by ultrasound on ice, centrifuging for 10min at 3000 r, taking the supernatant, and detecting the index level of triglyceride and the like in the supernatant. Meanwhile, liver tissue samples were subjected to histopathological analysis to evaluate the protective effect of compound HN-001 treatment on liver.
2. Experimental results:
as a result, referring to fig. 6 to 7, it can be seen from fig. 6 that intraperitoneal injection of the compound HN-001 into mice is effective in inhibiting weight gain in mice induced by long-term HFC diet. After 7 weeks of compound HN-001 administration, HN-001 mice lost about 19.8% of their weight compared to model control mice, indicating that compound HN-001 significantly improved mouse obesity. The HN-001 group mice had significantly reduced liver weight compared to the model control group mice, and the HN-001 group mice had an effective reduction in liver/body weight ratio.
As can be seen from fig. 7, HN-001 group significantly reduced triglyceride levels in liver tissue of mice compared to model control group mice. Blood biochemical analysis shows that HN-001 group significantly reduces the level of glutamic oxaloacetic transaminase and glutamic pyruvic transaminase in the blood of model mice compared with model control mice. The pathological staining result shows that the compound HN-001 can obviously protect liver cells, reduce liver lipid level and inhibit generation and accumulation of collagen fibers and myofibers of liver tissues.
EXAMPLE 6 toxicity of Marine Aspergillus source Compound HN-001 on liver cells
In this example, a test object of the compound HN-001 derived from Aspergillus sp.c1 was used to detect the toxic effect of HN-001 on liver cells.
1. The experimental method comprises the following steps:
HepG2 cells in logarithmic growth phase were taken according to 5X 10 3 Cells/well, plated evenly into 96-well plates, and cultured overnight. Diluting compound HN-001 with different concentrations of working solution (0.1, 0.5, 1, 5, 10, 25, 50, 100 μm), adding corresponding group of medicinal working solution, 100 μl/well, 37 ℃ and 5% CO into each well of 96-well plate 2 And (5) standing and culturing for 24 hours. ExperimentA control group, DMSO control group and compound HN-001-treated group were set. After 24 hours of completion of the action, the cell viability was examined by CCK-8 method.
The specific method for detecting the cell survival rate by the CCK-8 method comprises the following steps: after 24 hours of compound action, the medium was discarded, and 100. Mu.L of cell culture medium containing 10% CCK-8 action solution, 37℃and 5% CO was added to each well 2 The culture was allowed to stand for 4 hours, and the absorbance (OD) at 450nm was measured by an ELISA reader.
Wherein, the cell viability assay uses a vehicle (DMSO) control group as a control (Ctrl), and the fold relationship is:
the experimental results are the average of three independent experiments, and the results are statistically analyzed according to the average value + -standard deviation.
3. Experimental results:
as a result, referring to fig. 8, incubation of hepatocytes with compound HN-001 for 24 hours showed no inhibitory activity on cell viability and high safety, compared to control cells.
In conclusion, through in vivo and in vitro experiments, the compound HN-001 can effectively inhibit palmitic acid induced hepatotoxicity and effectively protect the liver in vivo, so that the nonalcoholic fatty liver can be effectively improved or treated, and the compound has low toxicity.
The above examples are preferred embodiments of the present invention, but the embodiments of the present invention are not limited to the above examples, and any other changes, modifications, substitutions, combinations, and simplifications that do not depart from the spirit and principle of the present invention should be made in the equivalent manner, and the embodiments are included in the protection scope of the present invention.
Claims (4)
1. Use of a marine aspergillus source compound in the preparation of a medicament for the treatment of non-alcoholic fatty liver disease, characterized in that the marine aspergillus source compound has the structure of formula (I):
2. the use according to claim 1, wherein said treatment of non-alcoholic fatty liver disease comprises inhibition of liver lipid accumulation, liver injury, liver inflammation and fibrosis.
3. The use according to claim 1, wherein the marine aspergillus source compound is also a pharmaceutically acceptable salt thereof.
4. The use according to any one of claims 1 to 3, wherein the medicament for treating non-alcoholic fatty liver disease is in the form of an oral or injectable preparation.
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| CN202210648711.4A Active CN115040502B (en) | 2022-06-09 | 2022-06-09 | Application of marine aspergillus source compound |
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Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US6087540A (en) * | 1996-04-22 | 2000-07-11 | Shionogi & Co., Ltd. | Terphenyl compounds and medicines containing the same |
| JP2003306409A (en) * | 2003-05-27 | 2003-10-28 | Kanebo Ltd | Terphenyl derivative and application |
| CN102079735A (en) * | 2010-10-22 | 2011-06-01 | 中山大学 | Terphenyl compound and preparation method thereof and application of compound as acetylcholinesterase inhibitor |
| CN107812182A (en) * | 2017-11-27 | 2018-03-20 | 中国药科大学 | The application of neuraminidase and inhibitor in the medicine for suppressing liver gluconeogenesis is prepared |
| WO2018160772A1 (en) * | 2017-02-28 | 2018-09-07 | The United State Of America, As Represented By The Secretary, Department Of Health & Human Services | Method of treating obesity, insulin resistance, non-alcoholic fatty liver disease including non-alcoholic steatohepatitis |
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2022
- 2022-06-09 CN CN202210648711.4A patent/CN115040502B/en active Active
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US6087540A (en) * | 1996-04-22 | 2000-07-11 | Shionogi & Co., Ltd. | Terphenyl compounds and medicines containing the same |
| JP2003306409A (en) * | 2003-05-27 | 2003-10-28 | Kanebo Ltd | Terphenyl derivative and application |
| CN102079735A (en) * | 2010-10-22 | 2011-06-01 | 中山大学 | Terphenyl compound and preparation method thereof and application of compound as acetylcholinesterase inhibitor |
| WO2018160772A1 (en) * | 2017-02-28 | 2018-09-07 | The United State Of America, As Represented By The Secretary, Department Of Health & Human Services | Method of treating obesity, insulin resistance, non-alcoholic fatty liver disease including non-alcoholic steatohepatitis |
| CN107812182A (en) * | 2017-11-27 | 2018-03-20 | 中国药科大学 | The application of neuraminidase and inhibitor in the medicine for suppressing liver gluconeogenesis is prepared |
Non-Patent Citations (1)
| Title |
|---|
| Zhi Kai Guo et al.."p-Terphenyl and Diterpenoid Metabolites from Endophytic Aspergillus sp.YXf3".Journal of Natural Products.2011,(第75期),第15-21页. * |
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