Application of andrographis paniculata diterpenoid compound in preparation of FABP4 inhibitor and osteoarthritis related drugs

Abstract

The invention relates to application of andrographis paniculata diterpenoid compounds in preparation of FABP4 inhibitors and osteoarthritis related drugs. The research finds that the andrographis paniculata diterpenoid compound can be used as an effective FABP4 inhibitor, can reduce ADAMTS4 overexpression in cartilage cells, and can be used for preparing the anti-osteoarthritis medicine. After normative, scientific and rigorous tests are carried out, the application of the andrographis paniculata diterpenoid compound, the pharmaceutically acceptable salt of the andrographis paniculata diterpenoid compound, the solvate of the andrographis paniculata diterpenoid compound or the andrographis paniculata diterpenoid compound derivative in preparation of the FABP4 inhibitor is provided. Experiments prove that the andrographis paniculata diterpenoid compound can be used as an effective FABP4 inhibitor, can inhibit ADAMTS4 overexpression induced by FABP4, relieve hose degradation and relieve cartilage injury and / or cartilage erosion caused by osteoarthritis, and can be used for preparing the anti-osteoarthritis medicine.

Description

Technical Field

[0001] The present invention relates to the field of medicines, and in particular to the application of andrographolide diterpenoid compounds in the preparation of FABP4 inhibitors and osteoarthritis related drugs. Background Art

[0002] Osteoarthritis (OA) is a common degenerative joint disease characterized by the progressive deterioration of ligaments and the breakdown of cartilage. The condition involves the gradual erosion of joint cartilage, leading to pain, stiffness, and limited movement. As cartilage degenerates, the bones within the joint begin to rub against each other, causing additional damage and discomfort. Therefore, hindering cartilage degradation is a promising treatment approach for developing OA drugs.

[0003] Recent studies have shown that FABP4 contributes to the degradation of cartilage. FABP4 is a cytoplasmic protein with a molecular weight of approximately 15 kDa that promotes the transfer of fatty acids between the extracellular and intracellular membranes. Epidemiological surveys have shown that FABP4 expression is elevated in the synovial tissue of OA patients, and the level of FABP4 in the synovial fluid is positively correlated with the severity of cartilage degeneration. Small molecule inhibitors of FABP4 and knockout of FABP4 have been shown to improve cartilage damage in OA animal models. Therefore, FABP4 has become a key factor in the induction of OA. However, existing small molecule inhibitors have limited inhibitory effects on FABP4 and have little effect on alleviating the symptoms associated with osteoarthritis. Summary of the invention

[0004] This study found that andrographolide diterpenoids can be used as effective FABP4 inhibitors to reduce ADAMTS4 overexpression in chondrocytes and can be used to prepare anti-osteoarthritis drugs. After conducting standardized, scientific and rigorous experiments, the application of andrographolide diterpenoids, pharmaceutically acceptable salts of andrographolide diterpenoids, solvates of andrographolide diterpenoids or andrographolide diterpenoid derivatives in the preparation of FABP4 inhibitors is provided.

[0005] Experimental results have shown that andrographolide diterpenoids can serve as effective FABP4 inhibitors, inhibit FABP4-induced ADAMTS4 overexpression, alleviate cartilage degradation, and reduce cartilage damage and / or cartilage erosion in osteoarthritis, and can be used to prepare anti-osteoarthritis drugs.

[0006] In one embodiment, the andrographolide diterpenoid compound includes andrographolide.

[0007] A FABP4 inhibitor includes andrographolide diterpenoid compounds, pharmaceutically acceptable salts of andrographolide diterpenoid compounds, solvates of andrographolide diterpenoid compounds or andrographolide diterpenoid compound derivatives.

[0008] In one embodiment, the andrographolide compound is andrographolide.

[0009] In one embodiment, the IC50 value of the andrographolide diterpenoid compound is 4.61 μM.

[0010] In one embodiment, the andrographolide diterpenoid compound or the above-mentioned FABP4 inhibitor is used in the preparation of a drug for reducing the overexpression of ADAMTS4 in chondrocytes.

[0011] The ADAMTS4 overexpression is ADAMTS4 overexpression caused by FABP4.

[0012] Use of andrographolide diterpenoid compounds or the above-mentioned FABP4 inhibitors in the preparation of anti-osteoarthritis drugs.

[0013] In one embodiment, the anti-osteoarthritis drug can reduce degradation of the hose.

[0014] In one embodiment, the anti-osteoarthritis drug can reduce cartilage damage and / or cartilage erosion caused by osteoarthritis. BRIEF DESCRIPTION OF THE DRAWINGS

[0015] Figure 1 This is a diagram showing the detection results of the effect of FABP4 on the expression of ADAMTS4;

[0016] Figure 2 In the figure, A is the chemical structure of diterpenoids of andrographolide; B is the molecular docking model; C is the dose-response curve of andrographolide, deoxyandrographolide, isoandrographolide and BMS309403;

[0017] Figure 3 In the figure, A and B are test results of the effect of BMS309403 on ADAMTS4 overexpression at the protein and mRNA levels; C and D are test results of the effect of AP on ADAMTS4 overexpression at the protein and mRNA levels;

[0018] Figure 4 In the figure, A is a flowchart of the animal study plan; B is an image of HE, crocin O, and toluidine blue staining of the slices (scale bar: 100 μm); cartilage was stained orange-red and blue with crocin O and toluidine blue, respectively; the arthritis index (see C) and score (see D) were calculated to evaluate the anti-OA activity of FABP4 inhibitors; E is the collection of fluid for IL-1β, TNF-α, and ADAMTS-4 quantification. The results were statistically analyzed using the t test;

[0019] Figure 5Immunohistochemical staining of FABP4 (A) and ADAMTS4 (C) in cartilage; quantitative analysis of FABP4 (B) and ADAMTS4 (D) positive cells among different groups (n=6);

[0020] Figure 6 In the figure, A is a microscopic CT scan image of the knee joint showing bone regeneration of knee cartilage after administration of andrographolide and BMS309403; B to E are the quantitative detection results of andrographolide and BMS3093403 on bone regeneration of knee cartilage. DETAILED DESCRIPTION

[0021] In order to make the above-mentioned objects, features and advantages of the present invention more obvious and easy to understand, the specific implementation of the present invention is described in detail below in conjunction with specific embodiments and drawings. In the following description, many specific details are set forth to facilitate a full understanding of the present invention. However, the present invention can be implemented in many other ways different from those described herein, and those skilled in the art can make similar improvements without violating the connotation of the present invention, so the present invention is not limited by the specific implementation disclosed below.

[0022] One embodiment of the present study provides the use of andrographolide compounds, pharmaceutically acceptable salts of andrographolide compounds, solvates of andrographolide compounds, or andrographolide derivatives in the preparation of FABP4 inhibitors.

[0023] Fatty acid-binding protein 4 (FABP4) is a member of the intracellular lipid-binding protein superfamily. It exists in the cells of vertebrates and invertebrates and plays an important role in the uptake, transport and metabolic regulation of long-chain fatty acids, and is involved in other life processes of organisms. The role of FABP4 in cells is to promote fatty acid solubilization, transport and metabolism. The exact mechanism by which FABP4 participates in the pathogenesis of OA is still unclear. One reasonable explanation is that FABP4 promotes the transport of saturated fatty acids and causes cartilage damage by upregulating metalloproteinases in chondrocytes. In animal models of OA, BMS309403, as a FABP4 inhibitor, has been shown to alleviate symptoms by reducing cartilage degradation and inhibiting the expression of metalloproteinases. These findings suggest that small molecule targeted inhibition of FABP4 may be an important strategy for the treatment of OA. In addition to synthetic compounds, natural products are another potential source for the discovery of FABP4 inhibitors.

[0024] This study found that Andrographis paniculata extract was shown to protect OA patients from cartilage damage and pain. Experimental verification showed that Andrographis paniculata diterpenoids can be used as effective FABP4 inhibitors, inhibit FABP4-induced ADAMTS4 overexpression, alleviate cartilage degradation, and reduce cartilage damage and / or cartilage erosion in osteoarthritis, and can be used to prepare anti-osteoarthritis drugs.

[0025] In one embodiment, the diterpenoids of Andrographis paniculata include andrographolide. Andrographolide (AP) is the main diterpenoid in Andrographis paniculata, the main bioactive component of Andrographis paniculata extract, and has been shown to reduce the level of metalloproteinases in chondrocytes. The IC50 value of the diterpenoids of Andrographis paniculata is 4.61 μM, and the inhibitory effect on FABP4 is very obvious.

[0026] It should be noted that andrographolide compounds include andrographolide, deoxyandrographolide, isoandrographolide, etc.

[0027] One embodiment of the present study also provides a FABP4 inhibitor, including andrographolide diterpenoid compounds, pharmaceutically acceptable salts of andrographolide diterpenoid compounds, solvates of andrographolide diterpenoid compounds, or andrographolide diterpenoid compound derivatives.

[0028] Experimental results have shown that andrographolide diterpenoids can serve as effective FABP4 inhibitors, inhibit FABP4-induced ADAMTS4 overexpression, alleviate cartilage degradation, and reduce cartilage damage and / or cartilage erosion in osteoarthritis, and can be used to prepare anti-osteoarthritis drugs.

[0029] In one embodiment, the andrographolide compound is andrographolide. The IC50 value of the andrographolide compound is 4.61 μM, and the inhibitory effect on FABP4 is very obvious. IC50 generally refers to the half inhibitory concentration.

[0030] It should be noted that andrographolide compounds include andrographolide, deoxyandrographolide, isoandrographolide, etc.

[0031] One embodiment of the present study also provides the use of andrographolide diterpenoid compounds or the above-mentioned FABP4 inhibitors in the preparation of drugs for reducing the overexpression of ADAMTS4 in chondrocytes.

[0032] ADAMTS4 is a degradative protein and metalloproteinase 4. This study found that FABP4 dose-dependently increased degradative protein and metalloproteinase 4 (ADAMTS4), but had no significant effect on the expression of matrix metalloproteinase 3 (MMP3) and matrix metalloproteinase 13 (MMP13). Andrographis paniculata diterpenoids can reduce the overexpression of ADAMTS4 in chondrocytes and can be used to prepare drugs that reduce the overexpression of ADAMTS4 in chondrocytes.

[0033] One embodiment of the present study also provides the use of andrographolide diterpenoid compounds or the above-mentioned FABP4 inhibitors in the preparation of anti-osteoarthritis drugs.

[0034] This study found that andrographolide diterpenoid compounds can alleviate the ability of cartilage degradation in patients with osteoarthritis, reduce cartilage damage and / or cartilage erosion in osteoarthritis, play a protective role on chondrocytes, and can be used to prepare anti-osteoarthritis drugs.

[0035] The following are specific embodiments.

[0036] Unless otherwise specified, the drugs and instruments used in the examples are all conventionally selected in the art. Experimental methods without specific conditions in the examples are usually carried out under conventional conditions, such as those described in literature or books or methods recommended by kit manufacturers.

[0037] Example 1 Materials and Methods

[0038] 1. Molecular docking screening

[0039] For molecular docking, the FABP4 structure file was obtained from the protein data bank (PDB3FR5), and the database was docked with FABP4 using AutoDock Vinav.1.0.2 (The Scripps Research Institute, CA). The docking results were then visualized using Pymol 1.3 (DeLano Scientific, San Carlos, CA).

[0040] 2. Cell culture and reagents

[0041] Human chondrocytes C28 / I2 were cultured in DMEM (Sigma-Aldrich, St. Louis, MO) supplemented with 10% fetal bovine serum (Hyclone), 100 units / mL penicillin and 100 μg / mL streptomycin. All cell culture reagents were from Intron (Carlsbad, CA). FABP4 inhibitor AP was from the National Institute of Food and Drug Control (Carlsbad, CA), and BMS309403 was from Cayman Chemical Company (Ann Arbor, MI, USA).

[0042] 3. Western Blotting

[0043] To analyze the expression of ADAMTS4, C28 / I2 cells were seeded in 12-well plates and treated with the desired concentrations of FABP4, FABP4, and FABP4 inhibitors for 48 h. After cell lysis, the proteins were purified and subjected to Western blot analysis. In the phosphorylation experiment, human chondrocytes C28 / I2 were stimulated with FABP4, cultured in the presence of AP for 48 h, and the cell lysates were collected for immunoblot analysis to quantify the phosphorylation of NF-κB. The proteins were separated by electrophoresis on 10% SDS polyacrylamide gels, and the transferred proteins were detected with primary antibodies against ADAMTS4, p-IKK, pp-p65, IKK, p65, or GAPDH (Bioss, China) for 18 h. The cell membranes were then washed and incubated with horseradish peroxidase-conjugated secondary antibodies for 1 h. The blots were visualized using chemiluminescent detection reagents (Millipore) and imaged with a gel Doc imager (Bio-Rad, CA, US).

[0044] 4. RT-qPCR analysis

[0045] C28 / I2 cells were plated in 12-well plates and treated with the desired concentrations of FABP4 and FABP4 inhibitors for 24 h at 37°C. Next, cells were lysed using TRIZOL reagent (Takara) and reverse transcribed using PrimeScript RT Master Mix Kit (Takara). RT-qPCR was performed on a LightCycler96 instrument.

[0046] 5. RNA sequencing and data analysis

[0047] C28 / I2 cells were seeded in 6-well plates and incubated for 24 h before the addition of FABP4. After treatment, cells were collected and sent to the BGI Genomics Facility for RNA extraction and bulk mRNA sequencing. Library preparation was performed and paired-end sequencing was performed on an Illumina HiSeq2000 sequencer (Illumina, USA). SOAPnuke (v1.5.2) was used to perform quality control on raw sequencing reads (fastq), and Bowtie2 (v2.2.5) was used to align the resulting clean reads to the genome. RSEM (v1.2.12) was used to detect the expression levels of genes, and DESeq2 (v1.4.5) was used to analyze differential expression levels.

[0048] 6. FABP4 fluorescence displacement assay

[0049] 1,8-ANS (10 μM, Sigma, St. Louis, MO) was mixed with FABP4 protein (10 μM, final concentration) in phosphate buffered saline (PBS, pH 7.4). The desired concentration of the compound was then added and incubated for 3 minutes at room temperature. The fluorescence signal at 370 nm (excitation) and 470 nm (emission) was measured by an i3 multi-mode microplate reader (Molecular Devices, CA, USA).

[0050] 7. Animal model design

[0051] The rats used were female Wistar rats aged 12-16 weeks, housed in the SIAT Animal Breeding Center under controlled temperature conditions. The rats were divided into 6 groups, each with 10 rats. One group served as a blank control for sham surgery experiments, and the other five groups served as OA model experiments. The negative control group was treated with saline, the positive control group with BMS309403 15 mg / kg / day, the AP low-dose group 5 mg / kg / day, the AP medium-dose group 15 mg / kg / day, and the AP high-dose group 30 mg / kg / day. These drugs were administered by intraperitoneal injection for 4 weeks. After the treatment period, all rats were euthanized using overdose of anesthesia, and the joints were collected for further analysis of their histological and morphological characteristics. This study was approved by the Institutional Animal Care and Use Committee (IACUC) at SIAT with the registration number SIAT-IACUC-190711-YGS-WY-A0774.

[0052] 8. Micro-CT

[0053] For micro-CT imaging of rat knee joints, the joints were first fixed with 4% neutral formalin for 2 days. Then, they were placed in a 34 mm scanning tube and scanned using a micro-CT machine (Scanco Microct, μCT100, Switzerland) with a resolution of 18 μm and a voltage / current value of 55 kV / 200 μA. The images and reconstruction data were obtained and analyzed using CTan and CTvol software from Bruker (Belgium).

[0054] 9. Histological analysis

[0055] Then decalcify with 10% EDTA at pH 7.4 for 30 days, embed in paraffin, and cut into 5 μm thick slices. The slices were dewaxed with xylene, rehydrated with graded ethanol, stained with hematoxylin and eosin (H&E), crocin O and toluidine blue (Sigma, US) (Sigma, US) for tissue visualization. The expression levels of ADAMTS4 and FABP4 were detected with corresponding antibodies. Finally, the slices were sealed with neutral glue and imaged using an automatic slide scanner (SLIDEVIEW VS200, OLYMPUS, Japan).

[0056] 10.ELISA experiment

[0057] Fluids from euthanized rats, inflammatory factors IL-1β (IL-1β ELISA Kit, PI301, Beyotime Biotech, Shanghai, China), TNF-α (TNF-α ELISA Kit, PT512, Beyotime Biotech, Shanghai, China), and ADAMTS4 (Mouse ADAMTS4 ELISA kit, CSB-EL001311MO, CUSABIO, Wuhan, China) were detected using ELISA kits according to the manufacturer’s instructions.

[0058] 11. Cytotoxicity

[0059] The cells were seeded in a 96-well plate and treated with the drug solution at the desired concentration. After incubation for 48 hours, 10% cell counting kit-8 (CCK8) solution (Dojindo, Kumamoto, Japan) was added and incubated for 15 minutes. The absorbance was measured at 450 nm using an ELISA reader.

[0060] 12. Statistics

[0061] Student's t-test was performed using GraphPad software (CA, US) to determine statistical significance. Results are presented as mean ± SEM and were considered significant when p value was < 0.05.

[0062] Example 2

[0063] FABP4 can upregulate the expression of ADAMTS4

[0064] In order to study the effect of FABP4 on chondrocytes, chondrocytes were treated with FABP4 at different concentrations (0, 6.25 ng / mL, 12.5 ng / mL, 25 ng / mL), and then detected by Western blotting. The specific detection process is detailed in Example 1. The detection results are detailed in Figure 1 . Figure 1 This is a graph showing the detection results of the effect of FABP4 on ADAMTS4 expression.

[0065] Example 3

[0066] Andrographolide (AP) was identified as a FABP4 inhibitor

[0067] The molecular docking screening experimental method of Example 1 was used to identify the diterpenoid compounds of Andrographis paniculata, and the dose-response curves of inhibitors (andrographolide, deoxyandrographolide, isoandrographolide and BMS309403) were obtained by FABP4 activity inhibition test. The test results are detailed in Figure 2 . Figure 2 In the figure, A is the chemical structure of andrographolide diterpenoids, B is the molecular docking model, and C is the dose-response curve of inhibitors (andrographolide, deoxyandrographolide, and isoandrographolide) and BMS309403.

[0068] like Figure 2 As shown, andrographolide, deoxyandrographolide, and isoandrographolide have the lowest binding energy to FABP4 (PDB code 3FR5). The three-dimensional plot depicts the interaction of the ligands (color sticks) with human FABP4 (yellow cartoon) with the key residues labeled. Andrographolide (AP) is the strongest inhibitor of FABP4 among the andrographolide diterpenes. In the simulation, AP forms hydrogen bonds with Tyr-19, Ser-53, Ser-55, and Arg-78 in the FABP4 inhibitor binding pocket. Arg-78 has been reported as a key residue for FABP4 inhibitor binding, and deoxyandrographolide and isoandrographolide do not bind directly to Arg-78, which may explain why they have lower binding affinity to AP. In the fluorescence displacement assay, AP is the strongest FABP4 inhibitor with an IC50 value of 4.61 μM, while deoxyandrographolide and isoandrographolide have IC50 values ​​of 9.6 μM and 10.2 μM, respectively. Using the known FABP4 inhibitor BMS309403 as a positive control, it was found to be 5 times more potent than AP with an IC50 value of 0.9 μM.

[0069] Example 4

[0070] AP inhibits FABP4-induced ADAMTS4 overexpression

[0071] The materials and methods of Example 1 were used to detect the effects of BMS309403 and AP on ADAMTS4 overexpression at the protein and mRNA levels. The test results are detailed in Figure 3 . Figure 3 Middle A and Figure 3 Figure B shows the test results of the effect of BMS309403 on ADAMTS4 overexpression at the protein and mRNA levels. Figure 3 Middle C and Figure 3 D in the middle is the detection result of the effect of AP on ADAMTS4 overexpression at the protein and mRNA levels.

[0072] like Figure 3 Middle A and Figure 3 As shown in Figure B, BMS309403 reduced ADAMTS4 overexpression at both the protein and mRNA levels. Figure 3 As shown in Figure C, the inhibitory effect of AP on FABP4-induced ADAMTS4 upregulation was dose-dependent. Figure 3 As shown in D, quantitative PCR also confirmed the reduction of ADAMTS4 by AP. In addition, BMS309403 and AP did not change cell viability at the treatment dose.

[0073] Example 5

[0074] AP can alleviate the symptoms of MIA-induced OA in rat knee joints

[0075] The materials and methods of this example are shown in Example 1, and the test results are shown in Figure 4 and Figure 5 .

[0076] Figure 4 In the figure, A is a flowchart of the animal study plan; B is an image of HE, crocin O and toluidine blue staining of the slice (scale bar: 100 μm); cartilage was stained orange-red and blue with crocin O and toluidine blue, respectively; the arthritis index (see C) and score (see D) were calculated to evaluate the anti-OA activity of FABP4 inhibitors; E is the collection of fluid for IL-1β, TNF-α and ADAMTS-4 quantification. The results were statistically analyzed by t test. #p<0.05, ##p<0.01 compared with the sham operation group; *p<0.05, **p<0.01 compared with the OA model group.

[0077] Figure 5In the MIA-induced OA model, andrographolide significantly reduced the expression of ADAMTS4, but not FABP4. Immunohistochemical staining of FABP4 (A) and ADAMTS4 (C) in cartilage. Quantitative analysis of FABP4 (B) and ADAMTS4 (D) positive cells among different groups (n=6). ####p<0.001 compared with the control group; ****p<0.001 compared with the OA model group. Scale bar: 20 μm.

[0078] This example further evaluated the protective effect of AP on damaged joints using a mia-induced rat model. This model is widely used in OA drug development and is characterized by cartilage degradation. In addition, elevated levels of FABP4 in the joints were observed in mia-induced rat joints, which is consistent with human OA symptoms.

[0079] After the OA model was established by injection of iodoacetic acid, AP, BMS309403 or saline (sham group) were intraperitoneally injected daily for 4 weeks (see Figure 4 A), The body weight of the animals was monitored daily throughout the treatment period, and no significant weight loss was observed in the AP-treated group. After 4 weeks of treatment, the rats were euthanized, and their knee joints were collected for microtomography and histological examination for further analysis.

[0080] HE, crocin O, and toluidine blue staining showed that the surface of the articular cartilage in the sham operation group was smooth, clear, and intact, while the surface of the articular cartilage in the OA model group was rough and broken (see Figure 4 B). The cartilage layer in the OA model group was almost invisible, indicating that the OA model was successfully established (see Figure 4 B). There were fewer cartilage surface fractures in the medium and high dose AP groups, especially in the high dose group, which had almost no obvious cartilage surface damage (see Figure 4 B). The cartilage surface damage in the positive control group was also less (see Figure 4 B). To quantitatively evaluate cartilage damage, we performed a cartilage repair score. The results showed that both BMS309403 and high-dose AP significantly reduced cartilage surface damage (see Figure 4 C).

[0081] The arthritis index is a widely used tool for quantitatively evaluating knee arthritis in rodent models. The arthritis index of MIA model rats was higher, while that of BMS309403 group and AP group was lower (see Figure 5D), indicating that FABP4 inhibitors can alleviate the symptoms of joint arthritis in this model. AP has been reported to have anti-inflammatory properties and can also alleviate arthritis symptoms. We collected local fluid to test whether AP can reduce inflammatory cytokines. ELISA results showed that although AP did not affect Il-1β levels, it slightly reduced TNF-α levels, while BMS309403 had little effect on either Il-1β or TNF-α levels (see Figure 4 E).

[0082] To determine whether AP can reduce ADAMTS4 levels, we performed ELISA. The results showed that MIA significantly increased local ADAMTS4 levels, and both BMS309403 and AP treatments effectively reduced ADAMTS4 levels (Table S5). In addition, immunohistochemical staining further confirmed that both BMS309403 and AP treatments significantly reduced the expression of ADAMTS4 induced by MIA injection ( Figure 5 a and 5B). In MIA rats, FABP4 was upregulated in joint tissues, as previously reported. Interestingly, neither AP nor BMS309403 significantly altered FABP4 expression in the joints of MIA rats ( Figure 5 c and 5D). These results suggest that the protective effects of AP and BMS309403 on cartilage may be due to their ability to reduce the expression of ADAMTS4 rather than directly regulating the expression of FABP4.

[0083] Example 6

[0084] Micro-CT scanning to monitor the changes of subchondral bone in the knee joint of OA models

[0085] The materials and methods of this example are shown in Example 1, and the test results are shown in Figure 6 . Figure 6 : Andrographolide significantly reduced subchondral erosion in the MIA-induced OA model. Figure 6 Figure A is a microscopic CT scan image of the knee joint showing the bone regeneration of the knee cartilage after administration of andrographolide and BMS309403. Quantitative detection results of andrographolide and BMS3093403 on the bone regeneration of the knee cartilage: BV / TV (Figure B), Tb.Pf (Figure C), Tb.Sp (Figure D) and Tb.Th (Figure E) values ​​of the sham operation group, OA model group, BMS309403 and andrographolide treatment groups (n=6). ##p<0.01, ####p<0.001 compared with the sham operation group; **<0.05, **p<0.01, ***p<0.005, ****p<0.001 compared with the OA model group.

[0086] The coronal three-dimensional reconstruction images of the knee joint showed that the femoral and tibial articular surfaces were severely damaged in the OA model ( Figure 6 A). However, after treatment with FABP4 inhibitors, the joint surface was significantly repaired ( Figure 6 A). Osteoporosis and significant loss of bone volume are common in OA, as is the bone volume / tissue volume ratio (BV / TV, Figure 6 B), trabecular pattern factor (Tb.Pf, Figure 6 C), trabecular separation (Tb.Sp, Figure 6 D) and trabecular thickness (Tb.Th, Figure 6 E) There was significant improvement after administration of AP and BMS309403, suggesting that high-dose AP treatment can reduce cartilage damage and bone erosion in osteoarthritis animals.

[0087] In summary, this study found that andrographolide diterpenoids can serve as effective FABP4 inhibitors, inhibit ADAMTS4 overexpression, alleviate the ability of cartilage degradation in patients with osteoarthritis, reduce cartilage damage and / or cartilage erosion in osteoarthritis, play a protective role on chondrocytes, and can be used to prepare anti-osteoarthritis drugs.

[0088] The technical features of the above-described embodiments may be arbitrarily combined. To make the description concise, not all possible combinations of the technical features in the above-described embodiments are described. However, as long as there is no contradiction in the combination of these technical features, they should be considered to be within the scope of this specification.

[0089] The above-mentioned embodiments only express several implementation methods of the present invention, and the descriptions thereof are relatively specific and detailed, but they cannot be understood as limiting the scope of the invention patent. It should be pointed out that, for ordinary technicians in this field, several variations and improvements can be made without departing from the concept of the present invention, and these all belong to the protection scope of the present invention. Therefore, the protection scope of the patent of the present invention shall be subject to the attached claims.

Claims

1. Use of andrographolide compounds, pharmaceutically acceptable salts of andrographolide compounds, solvates of andrographolide compounds or derivatives of andrographolide compounds in the preparation of FABP4 inhibitors.

2. The use according to claim 1, characterized in that: The andrographolide compounds include andrographolide.

3. A FABP4 inhibitor, characterized in that The invention comprises andrographolide compounds, pharmaceutically acceptable salts of andrographolide compounds, solvates of andrographolide compounds or derivatives of andrographolide compounds.

4. The FABP4 inhibitor according to claim 3, characterized in that The andrographolide compound is andrographolide.

5. The FABP4 inhibitor according to claim 3, characterized in that The IC50 value of the andrographolide diterpenoid compound is 4.61 μM.

6. Use of an Andrographis paniculata diterpenoid compound or the FABP4 inhibitor according to any one of claims 3 to 5 in the preparation of a drug for reducing overexpression of ADAMTS4 in chondrocytes.

7. The use according to claim 6, characterized in that: The ADAMTS4 overexpression is ADAMTS4 overexpression caused by FABP4.

8. Use of an Andrographis paniculata diterpenoid compound or the FABP4 inhibitor according to any one of claims 3 to 6 in the preparation of an anti-osteoarthritis drug.

9. The use according to claim 8, characterized in that: The anti-osteoarthritis drug can alleviate the degradation of the hose.

10. The use according to claim 8, characterized in that: The anti-osteoarthritis drug can reduce cartilage damage and / or cartilage erosion caused by osteoarthritis.

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