Application of obesity factor AdipoQ receptor stimulant in treatment of obesity-related thyroid cancer

Abstract

The invention relates to the technical field of thyroid cancer treatment, in particular to application of an obesity factor AdipoQ receptor stimulant in treatment of obesity-related thyroid cancer. The invention provides a stable and reliable preparation method of AdipoQ, which can effectively improve the sensitivity of insulin and reduce the accumulation of fat in the body, and provides a functional verification method of an obesity factor AdipoQ receptor stimulant AdipoRon in treatment of obesity-related thyroid cancer for providing a feasible verification scheme and theoretical basis for subsequent clinical transformation. Meanwhile, the invention provides a pharmaceutical composition for treating obesity-related thyroid cancer, which can effectively inhibit the growth and migration ability of thyroid cancer cells, promote the apoptosis of thyroid cancer cells and effectively treat obesity-related thyroid cancer patients.

Description

Technical Field

[0001] The present invention relates to the technical field of thyroid cancer treatment, and particularly relates to the application of an adiponectin AdipoQ receptor agonist in the treatment of obesity-related thyroid cancer. Background Art

[0002] Obesity is a major health and social problem, greatly increasing the risk of various serious complications including the development of endocrine cancers. In recent years, obesity and thyroid cancer have been two hot topics in the endocrine field. There is sufficient evidence that obesity is a risk factor for the incidence of thirteen malignant tumors including thyroid cancer. Although most thyroid cancers have a low degree of malignant invasion, existing treatment methods are effective, and the long-term prognosis is good, it is still one of the malignant tumors with the fastest increasing incidence in China in recent years, and the mortality rate has also increased slightly. A small number of thyroid cancers with strong invasiveness quickly metastasize or recur, lacking effective treatment methods, which also seriously reduces the quality of life of patients.

[0003] In recent years, people's understanding of obesity-related malignancies has increased rapidly. Adipokines, as key factors affecting the pathophysiological processes of obesity-related cancers, have attracted more and more attention. AdipoQ has been considered a key mediator connecting obesity and endocrine-related malignancies, but the clinical translation of AdipoQ has been restricted by technical bottlenecks and cannot be carried out. As a receptor agonist of AdipoQ, AdipoRon can bind to and activate AdipoR1 and AdipoR2, improving insulin resistance and type 2 diabetes. It has the advantages of low synthesis cost and stable structure, but its application in thyroid cancer is still unknown.

[0004] In view of the above problems, the present invention provides a solution. Summary of the Invention

[0005] The purpose of the present invention is to provide the application of an adiponectin AdipoQ receptor agonist in the treatment of obesity-related thyroid cancer, which can effectively solve the application problem of the AdipoQ receptor agonist AdipoRon in the treatment of obesity-related thyroid cancer.

[0006] To achieve the above purpose, the present invention adopts the following technical scheme: An adiponectin AdipoQ, which is an endogenous bioactive polypeptide secreted by adipocytes (NBCI ID: KAL1288581.1), and its corresponding amino acid sequence is shown in SEQ ID NO: 1;

[0007] Furthermore, a preparation method of an adiponectin AdipoQ includes the following steps:

[0008] A1: Sterilize the ultraviolet disinfection operation table for 30 minutes. Subsequently, prepare the culture medium by mixing 44.5 ml of DMEM, 5 ml of FBS, and 0.5 ml of double antibiotics. Mix ampicillin, kanamycin, and streptomycin at a concentration of one-thousandth to make triple antibiotics. Take healthy human neck adipose tissue and soak it in 75% disinfected alcohol. After washing for 15 minutes, take out the human neck adipose tissue and transfer it to the triple antibiotics for soaking for 30 minutes;

[0009] A2: Transfer the soaked human neck adipose tissue to sterile water for soaking and washing for 15 minutes. Cut the adipose tissue into pieces and add 0.1% collagenase I. Digest at 37°C for 45 minutes, centrifuge at 1000 rpm for 5 minutes to obtain stromal vascular fragments. Put the stromal vascular fragments into the culture medium, and use a sterile glass slide to press the stromal vascular fragments against the bottom of the culture dish for normal culture. Observe once a day and change the culture medium every two days, waiting for adipose stem cells to crawl out from the bottom of the culture dish;

[0010] A3: When the density of adipose stem cells reaches 100% after culture, perform contact growth for 2 days. Subsequently, prepare the adipose cell induction medium. After preparation, use the adipose cell medium to induce for 2 days, and then replace the adipose cell medium and repeat the induction for 2 days;

[0011] A4: Resuscitate 293T cells, and use the culture medium of 44.5 ml of DMEM + 5 ml of FBS + 0.5 ml of double antibiotics medium. After the 293T cells are cultured to a density of 85%, rinse them with PBS, digest with trypsin for 2 minutes, and use the medium containing FBS to stop trypsin digestion;

[0012] A5: Aspirate and transfer the 293T cell suspension into a 15 ml centrifuge tube, add 6 ml of DMEM medium containing 10% FBS, balance and centrifuge at 1000 rpm / min for a total of 3 minutes. After centrifugation, discard the supernatant, add 1 ml of medium to resuspend the cells. Take 100 μL of the cell suspension, add 900 μL of PBS to resuspend the cells. Subsequently, take 10 μL of the cell suspension for cell counting. Take 2000 cells and add them to a 6-well plate containing 2 ml of medium and place them in a cell culture incubator with 5% CO2 for culture for 8 days;

[0013] A6: Construct a human AdipoQ recombinant plasmid pLV-EF1a-BSD-N-GFP-ADIPOQ using an empty vector, and then perform lentivirus packaging using a three-plasmid system. The tool cells are 293T cells that have completed resuscitation. Infect adipose stem cells with the virus solution and screen out AdipoQ adipose stem cells with stable expression. After the AdipoQ adipose stem cells with stable expression are cultured to a density of 85%, rinse them with PBS, digest with trypsin for 2 minutes, and terminate the trypsin digestion with a medium containing FBS. Then, aspirate and transfer the suspension of AdipoQ adipose stem cells with stable expression into a 15 ml centrifuge tube, add 6 ml of DMEM medium containing 10% FBS, balance and centrifuge at 1000 rpm / min for a total of 3 minutes;

[0014] A7: After centrifugation, discard the supernatant, add 1 ml of medium to resuspend the cells, evenly spread the resuspended cells in a 60 mm culture dish, shake the dish evenly in all directions, and then place the culture dish in a cell culture incubator with 5% CO2 for culture. Collect the medium of AdipoQ adipose stem cells with stable expression to obtain a medium containing the obesity factor AdipoQ. Balance and centrifuge the medium at 1000 rpm / min for a total of 3 minutes. After centrifugation, collect the supernatant to obtain a supernatant containing the obesity factor AdipoQ.

[0015] Further, the adipocyte induction medium described in step A3 is 44.5 ml of DMEM, 5 ml of FBS, 0.5 ml of double antibody, 0.5 mM 3-isobutyl-1-methylxanthine, 1.0 μM dexamethasone, 0.1 mM indomethacin, and 10 mg / L insulin; the empty vector described in step S6 is pLV-EF1a-BSD-N-GFP; the three-plasmid system described in step A6 is the pLV-EF1a-BSD-N-GFP-ADIPOQ, PVSVG, and PAX8 three-plasmid system;

[0016] Further, a pharmaceutical composition for treating obesity-related thyroid cancer uses the obesity factor AdipoQ receptor agonist AdipoRon as the active ingredient. In the pharmaceutical composition, the obesity factor AdipoQ receptor agonist AdipoRon accounts for 1-99% of the total weight of the drug, preferably 5-25%, and most preferably 10%;

[0017] The AdipoRon is Figure 1 the shown compound or the hydrate, solvate, metabolite, and pharmaceutically acceptable salt of the shown compound, with the molecular formula C 27 H 28 N2O3, a molecular weight of 428.52; a melting point of 106-108 °C, soluble in dimethyl sulfoxide, slightly soluble in water.

[0018] Furthermore, the pharmaceutical composition can be administered via the gastrointestinal tract or by inhalation. Preferably, the pharmaceutical dosage form for gastrointestinal administration is selected from: tablets, capsules, granules, solutions, extracts, pills, and powders.

[0019] Furthermore, a method for functional verification of the obesity factor AdipoQ receptor agonist AdipoRon in the treatment of obesity-related thyroid cancer includes the following steps:

[0020] B1: Use DMSO to prepare AdipoRon with gradient concentrations of oral activity to treat the differentiated thyroid cancer cell lines K-1 and KTC-1 for 24 hours. Apply the CCK8 cell cytotoxicity assay kit to detect the half-inhibitory concentration of thyroid cancer cells K-1 and KTC-1, namely IC50.

[0021] B2: Use the effective drug concentration corresponding to the IC50 value obtained in step B1 to test the effect of orally active AdipoRon on treating the thyroid cancer cell lines K-1 and KTC-1 for 0h, 24h, 48h, and 72h to test the effect of AdipoRon on the proliferation activity of the thyroid cancer cell lines.

[0022] B3: Use DMSO to prepare AdipoRon with gradient concentrations of oral activity for a plate colony formation assay. The experimental cells are K-1 and KTC-1. Subsequently, use the effective drug concentration corresponding to the IC50 value obtained in step B1 to treat K-1 and KTC-1 for 7 days to detect the effect of AdipoRon on the colony formation ability of thyroid cancer cells.

[0023] B4: Use a 20 μL pipette tip to scratch the cells in the densely populated areas of K-1 and KTC-1 cells with a cell concentration of 85%. Subsequently, use the effective drug concentration corresponding to the IC50 value obtained in step B1 to treat K-1 and KTC-1 cells for 24 hours to test the effect of AdipoRon on the migration ability of thyroid cancer cells.

[0024] B5: Use the effective drug concentration corresponding to the IC50 value obtained in step B1 to test the effect of orally active AdipoRon on treating the thyroid cancer cell lines K-1 and KTC-1 for 0h, 24h, 48h, and 72h. Subsequently, use the JC-1 fluorescent probe to detect the mitochondrial membrane potential of K-1 and KTC-1. When the mitochondrial membrane potential is high, JC-1 aggregates in the mitochondrial matrix to form polymers, which can produce red fluorescence. When the mitochondrial membrane potential is low, JC-1 cannot aggregate in the mitochondrial matrix, and at this time, JC-1 is a monomer, producing green fluorescence, to test the effect of AdipoRon on the apoptosis of thyroid cancer cells.

[0025] In summary, due to the adoption of the above technical solutions, the beneficial effects of the present invention are as follows: the present invention provides a stable and reliable method for preparing AdipoQ, which can effectively improve insulin sensitivity and reduce the accumulation of body fat. Moreover, the present invention provides a functional verification method for an obesity factor AdipoQ receptor agonist AdipoRon in the treatment of obesity-related thyroid cancer, providing a feasible verification scheme and theoretical basis for subsequent clinical transformation. At the same time, the present invention provides a pharmaceutical composition for treating obesity-related thyroid cancer, which can effectively inhibit the growth and migration ability of thyroid cancer cells, promote the apoptosis of thyroid cancer cells, and effectively treat patients with obesity-related thyroid cancer. BRIEF DESCRIPTION OF THE DRAWINGS

[0026] Figure 1 It is a molecular structure diagram of an obesity factor AdipoQ receptor agonist AdipoRon.

[0027] Figure 2 It is the half-maximal inhibitory concentration (IC50) of AdipoRon on thyroid cancer cell lines K-1 and KTC-1.

[0028] Figure 3 It is a diagram showing the effect of AdipoRon on thyroid cancer cell lines K-1 and KTC-1 detected by CCK8 at different time gradients.

[0029] Figure 4 It is a diagram showing the inhibitory effect of AdipoRon concentration gradient on the colony formation of K-1 and KTC-1.

[0030] Figure 5 It is a diagram showing the inhibitory effect of effective AdipoRon concentration on the colony formation of K-1 and KTC-1.

[0031] Figure 6 It is a diagram showing the inhibitory effect of AdipoRon on the migration ability of thyroid cancer cells.

[0032] Figure 7 It is a diagram showing the effect of AdipoRon on the apoptosis of thyroid cancer cells. DETAILED DESCRIPTION OF THE EMBODIMENTS

[0033] In order to make the technical means, creative features, achieved purposes and functions of the present invention easy to understand, the present invention will be further described below in conjunction with specific embodiments and diagrams.

[0034] The experimental methods in the following embodiments are all conventional methods unless otherwise specified, and are carried out according to the techniques or conditions described in the literature in this field or according to the product instructions. The materials, reagents, etc. used in the following embodiments can be obtained from commercial channels unless otherwise specified.

[0035] Example 1

[0036] 1: Sterilize the ultraviolet disinfection operating table for 30 minutes. Subsequently, prepare the culture medium: 44.5 ml of DMEM + 5 ml of FBS + 0.5 ml of double antibiotics. Mix ampicillin, kanamycin, and streptomycin with a concentration of one-thousandth to make triple antibiotics. Take healthy human neck adipose tissue and soak it in 75% disinfected alcohol. After washing for 15 minutes, take out the human neck adipose tissue and transfer it to the triple antibiotics for soaking for 30 minutes;

[0037] 2: Transfer the soaked human neck adipose tissue to sterile water for soaking and washing for 15 minutes. Cut the adipose tissue into pieces and add 0.1% collagenase I. Digest at 37°C for 45 minutes, centrifuge at 1000 rpm for 5 minutes to obtain stromal vascular fragments. Put the stromal vascular fragments into the culture medium, use a sterile glass slide to press the stromal vascular fragments on the bottom of the culture dish for normal culture. Observe once a day and change the culture medium every two days, waiting for adipose stem cells to crawl out from the bottom of the culture dish;

[0038] 3: When the density of adipose stem cells reaches 100% after culture, carry out contact growth for 2 days. Subsequently, prepare the adipose cell induction culture medium: 44.5 ml of DMEM, 5 ml of FBS, 0.5 ml of double antibiotics, 0.5 mM 3-isobutyl-1-methylxanthine, 1.0 μM dexamethasone, 0.1 mM indomethacin, and 10 mg / L insulin. After preparation, use the adipose cell culture medium to induce for 2 days, and then change the adipose cell culture medium and repeat the induction for 2 days;

[0039] 4: Resuscitate 293T cells, and use the culture medium of 44.5 ml of DMEM + 5 ml of FBS + 0.5 ml of double antibiotics. When the density of 293T cells reaches 85% after culture, rinse with PBS, digest with trypsin for 2 minutes, and use the culture medium containing FBS to stop the trypsin digestion;

[0040] 5: Aspirate and transfer the 293T cell suspension into a 15 ml centrifuge tube, add 6 ml of DMEM medium containing 10% FBS, balance and centrifuge at 1000 rpm / min for a total of 3 minutes. After centrifugation, discard the supernatant, add 1 ml of culture medium to resuspend the cells. Take 100 μL of the cell suspension, add 900 μL of PBS to resuspend the cells. Subsequently, take 10 μL of the cell suspension for cell counting. Take 2000 cells and add them to a 6-well plate containing 2 ml of culture medium and place them in a cell culture incubator with 5% CO2 for culturing for 8 days;

[0041] 6: The human AdipoQ recombinant plasmid pLV-EF1a-BSD-N-GFP-ADIPOQ was constructed using the empty vector pLV-EF1a-BSD-N-GFP. Subsequently, lentivirus packaging was performed using the three-plasmid system of pLV-EF1a-BSD-N-GFP-ADIPOQ, PVSVG, and PAX8. The tool cells used were 293T cells that had been resuscitated. Adipose stem cells were infected with the virus solution and stable-expressing AdipoQ adipose stem cells were screened out. After the stable-expressing AdipoQ adipose stem cells were cultured to a density of 85%, they were rinsed with PBS, digested with trypsin for 2 minutes, and the trypsin digestion was terminated using a medium containing FBS. Subsequently, the suspension of stable-expressing AdipoQ adipose stem cells was aspirated and loaded into a 15 ml centrifuge tube, 6 ml of DMEM medium containing 10% FBS was added, and it was balanced and centrifuged at 1000 rpm / min for a total of 3 minutes;

[0042] 7: After centrifugation, the supernatant was discarded, 1 ml of medium was added to resuspend the cells, and the resuspended cells were evenly spread in a 60 mm culture dish. The dish was shaken evenly up, down, left, and right, and then the culture dish was placed in a cell culture incubator with 5% CO2 for culture. The medium of stable-expressing AdipoQ adipose stem cells was collected to obtain the medium containing the obesity factor AdipoQ. The medium was balanced and centrifuged at 1000 rpm / min for a total of 3 minutes. After centrifugation, the supernatant was collected to obtain the supernatant containing the obesity factor AdipoQ prepared in Example 1.

[0043] Example 2

[0044] 1: Oral-active AdipoRon with gradient concentrations (AdipoRon concentrations were 10 μM, 20 μM, 30 μM, 40 μM, 50 μM in sequence) was configured using DMSO and used to treat the differentiated thyroid cancer cell lines K-1 and KTC-1 for 24 h. The CCK8 cell cytotoxicity assay kit was applied to detect the half-inhibitory concentration of thyroid cancer cells K-1 and KTC-1, that is, IC50. The results were as Figure 2 shown. It can be seen that the IC50 of thyroid cancer cells K-1 was 27.88 μM, and the IC50 of thyroid cancer cells KTC-1 was 28.84 μM;

[0045] 2: Thyroid cancer cell lines K-1 and KTC-1 were treated with oral-active AdipoRon at 27.88 μM and 28.84 μM respectively for 0 h, 24 h, 48 h, and 72 h to test the effect of AdipoRon on the proliferation activity of thyroid cancer cell lines. The results were as Figure 3 shown;

[0046] 3: Prepare serial dilutions of the orally active AdipoRon in DMSO for the colony formation assay. The experimental cells used were K-1 and KTC-1. The thyroid cancer cell lines K-1 and KTC-1 were treated with 27.88 μM and 28.84 μM of the orally active AdipoRon respectively for 7 days to examine the effect of AdipoRon on the colony formation ability of thyroid cancer cells. The results are shown in Figure 4 and Figure 5 as follows;

[0047] 4: Use a 20 μL pipette tip to create a cell scratch in the confluent areas of K-1 and KTC-1 cells with a cell density of 85%. Subsequently, the thyroid cancer cell lines K-1 and KTC-1 were treated with 27.88 μM and 28.84 μM of the orally active AdipoRon respectively for 24 h to test the effect of AdipoRon on the migration ability of thyroid cancer cells. The results are shown in Figure 6 as follows;

[0048] 5: Use the effective drug concentration corresponding to the IC50 value obtained in step B1 to treat the thyroid cancer cell lines K-1 and KTC-1 with the orally active AdipoRon for 0 h, 24 h, 48 h, and 72 h. Subsequently, use the JC-1 fluorescent probe to detect the mitochondrial membrane potential of K-1 and KTC-1. When the mitochondrial membrane potential is high, JC-1 aggregates in the mitochondrial matrix to form polymers, which can produce red fluorescence. When the mitochondrial membrane potential is low, JC-1 cannot aggregate in the mitochondrial matrix, and at this time JC-1 is a monomer, producing green fluorescence, to test the effect of AdipoRon on the apoptosis of thyroid cancer cells. The results are shown in Figure 7 as follows.

[0049] By analyzing Figures 2-7 it can be seen that AdipoRon has an inhibitory effect on the proliferation activity of thyroid cancer cells and shows time-dependence; AdipoRon significantly inhibits the colony formation ability of thyroid cancer cells; AdipoRon significantly inhibits the migration ability of thyroid cancer cells; after treatment with AdipoRon, the mitochondrial membrane potential of thyroid cancer cells decreases significantly, indicating that AdipoRon can significantly induce the apoptosis of thyroid cancer cells.

[0050] The above shows and describes the basic principles, main features, and advantages of the present invention. Those skilled in the art should understand that the present invention is not limited by the above embodiments. The above embodiments and the descriptions in the specification only illustrate the principles of the present invention. Without departing from the spirit and scope of the present invention, the present invention will have various changes and improvements, and these changes and improvements all fall within the scope of the present invention claimed. The scope of protection of the present invention is defined by the appended claims and their equivalents.

Claims

1. An obesity factor AdipoQ, characterized in that, The obesity factor AdipoQ is an endogenous bioactive polypeptide secreted by adipocytes, and its corresponding amino acid sequence is shown in SEQ ID NO:

1.

2. A preparation method of adiponectin, characterized in that, It includes the following steps: A1: Disinfect the operating table with ultraviolet light for 30 minutes. Subsequently, prepare the culture medium: 44.5 ml of DMEM + 5 ml of FBS + 0.5 ml of double antibiotics. Mix ampicillin, kanamycin, and streptomycin with a concentration of one-thousandth to make triple antibiotics. Soak healthy human neck adipose tissue in 75% disinfected alcohol, take out the human neck adipose tissue after washing for 15 min and transfer it to the triple antibiotics for soaking for 30 min; A2: Transfer the soaked human neck adipose tissue to sterile water for soaking and washing for 15 min. Cut the adipose tissue into pieces and add 0.1% collagenase I. Digest at 37 °C for 45 minutes, centrifuge at 1000 rpm for 5 minutes to obtain stromal vascular fragments. Put the stromal vascular fragments into the culture medium, use a sterile glass slide to press the stromal vascular fragments at the bottom of the culture dish for normal culture. Observe once a day and change the culture medium every two days, and wait for adipose stem cells to crawl out from the bottom of the culture dish; A3: When the density of adipose stem cells reaches 100% after culture, carry out contact growth for 2 days. Subsequently, prepare the adipose cell induction medium. After preparation, induce with the adipose cell medium for 2 days, and then change the adipose cell medium and repeat the induction for 2 days; A4: Resuscitate 293T cells, and use the culture medium of 44.5 ml of DMEM + 5 ml of FBS + 0.5 ml of double antibiotics. When the density of 293T cells reaches 85% after culture, rinse with PBS, digest with trypsin for 2 minutes, and use the culture medium containing FBS to stop trypsin digestion; A5: Pipette the 293T cell suspension into a 15 ml centrifuge tube, add 6 ml of DMEM medium containing 10% FBS, balance and centrifuge at 1000 rpm / min for a total of 3 minutes. After centrifugation, discard the supernatant, add 1 ml of culture medium to resuspend the cells. Take 100 μL of the cell suspension, add 900 μL of PBS to resuspend the cells, then take 10 μL of the cell suspension for cell counting. Take 2000 cells and add them to a 6-well plate containing 2 ml of culture medium and place them in a cell culture incubator with 5% CO2 for culture for 8 days; A6: Construct the human AdipoQ recombinant plasmid pLV-EF1a-BSD-N-GFP-ADIPOQ using an empty vector. Subsequently, carry out lentivirus packaging using the three-plasmid system. The tool cells are the resuscitated 293T cells. Infect adipose stem cells with the virus solution and screen out the AdipoQ adipose stem cells with stable expression. When the density of the AdipoQ adipose stem cells with stable expression reaches 85% after culture, rinse with PBS, digest with trypsin for 2 minutes, and use the culture medium containing FBS to stop trypsin digestion. Then pipette the AdipoQ adipose stem cell suspension with stable expression into a 15 ml centrifuge tube, add 6 ml of DMEM medium containing 10% FBS, balance and centrifuge at 1000 rpm / min for a total of 3 minutes; A7: After centrifugation, discard the supernatant, add 1 ml of culture medium to resuspend the cells, evenly spread the resuspended cells in a 60-mm culture dish, gently shake the dish up, down, left, and right, then place the culture dish in a cell incubator with 5% CO2 for culture. Collect the culture medium of AdipoQ adipose stem cells with stable expression to obtain the culture medium containing the obesity factor AdipoQ. Balance and centrifuge the culture medium at 1000 rpm / min for a total of 3 minutes. After centrifugation, collect the supernatant to obtain the supernatant containing the obesity factor AdipoQ.

3. The preparation method of an obesity factor AdipoQ according to claim 2, characterized in that, The adipocyte induction medium described in step A3 is composed of 44.5 ml of DMEM, 5 ml of FBS, 0.5 ml of penicillin-streptomycin, 0.5 mM of 3-isobutyl-1-methylxanthine, 1.0 μM of dexamethasone, 0.1 mM of indomethacin, and 10 mg / L of insulin.

4. The preparation method of an adiposity factor AdipoQ according to claim 2, characterized in that, The empty vector described in step A6 is pLV-EF1a-BSD-N-GFP; the three-plasmid system is the pLV-EF1a-BSD-N-GFP-ADIPOQ, PVSVG, and PAX8 three-plasmid system.

5. A functional verification method of an adipokine AdipoQ receptor agonist AdipoRon in the treatment of obesity-related thyroid cancer, characterized in that, It includes the following steps: B1: Use DMSO to prepare oral active AdipoRon with gradient concentrations to treat the differentiated thyroid cancer cell lines K-1 and KTC-1 for 24 h. Apply the CCK8 cytotoxicity assay kit to detect the half-inhibitory concentration of thyroid cancer cells K-1 and KTC-1, namely IC50. B2: Use the effective drug concentration corresponding to the IC50 value obtained in step B1 to test the oral active AdipoRon to treat the thyroid cancer cell lines K-1 and KTC-1 for 0 h, 24 h, 48 h, and 72 h to test the effect of AdipoRon on the proliferation activity of the thyroid cancer cell lines. B3: Use DMSO to prepare oral active AdipoRon with gradient concentrations for the plate colony formation assay. The experimental cells are K-1 and KTC-1. Then use the effective drug concentration corresponding to the IC50 value obtained in step B1 to treat K-1 and KTC-1 for 7 days to detect the effect of AdipoRon on the colony formation ability of thyroid cancer cells. B4: Use a 20-μL pipette tip to scratch the cells in the dense area of K-1 and KTC-1 cells with a cell concentration of 85%. Then use the effective drug concentration corresponding to the IC50 value obtained in step B1 to treat K-1 and KTC-1 cells for 24 h to test the effect of AdipoRon on the migration ability of thyroid cancer cells. B5: Use the effective drug concentration corresponding to the IC50 value obtained in step B1 to test the oral activity of AdipoRon on the thyroid cancer cell lines K-1 and KTC-1. The treatment time is 0 h, 24 h, 48 h, and 72 h. Subsequently, use the JC-1 fluorescent probe to detect the mitochondrial membrane potential of K-1 and KTC-1. When the mitochondrial membrane potential is high, JC-1 aggregates in the mitochondrial matrix to form polymers, which can produce red fluorescence. When the mitochondrial membrane potential is low, JC-1 cannot aggregate in the mitochondrial matrix. At this time, JC-1 is a monomer and produces green fluorescence, so as to test the effect of AdipoRon on the apoptosis of thyroid cancer cells.

6. A pharmaceutical composition for treating obesity-related thyroid cancer, characterized in that, The drug composition uses the adipokine AdipoQ receptor agonist AdipoRon as the active ingredient. In the drug composition, the adipokine AdipoQ receptor agonist AdipoRon accounts for 1-99% of the total weight of the drug, preferably 5-25%, and most preferably 10%.

7. A pharmaceutical composition for treating obesity-related thyroid cancer according to claim 6, characterized in that, The AdipoRon described above is the compound shown in Figure 1 or a hydrate, solvate, metabolite, and pharmaceutically acceptable salt of the shown compound, with the molecular formula C 27 H 28 N2O3, with a molecular weight of 428.52, a melting point of 106 - 108 °C, soluble in dimethyl sulfoxide and slightly soluble in water.

8. The pharmaceutical composition for treating obesity-related thyroid cancer according to claim 6, wherein, The drug composition can be administered via the gastrointestinal tract or by inhalation. Preferably, the dosage forms of the drug administered via the gastrointestinal tract are selected from: tablets, capsules, granules, solutions, instant granules, pills, and powders.

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