New medical application of ailanone

Abstract

The invention provides a novel medical application of ailanone. Specifically, the invention relates to a novel application of ailanone in preparation of an LSD1 protein degradation agent. The ailanthus altissima ketone is extracted from a medicinal plant ailanthus altissima, and can significantly degrade LSD1 protein in a dose-dependent and time-dependent manner. The invention also provides an application of ailanone as an LSD1 protein degradation agent in preparation of an LSD1 targeted drug. The invention also provides application of the LSD1 protein degradation agent in preparation of drugs for prevention and / or treatment of diseases related to abnormal LSD1 activity or expression. As an effective LSD1 protein degradation agent, ailanone has a relatively strong killing effect on tumor cells, can remarkably inhibit proliferation of various tumor cells, can inhibit growth of xenograft tumors of the tumor cells, and has relatively great potential for tumor treatment. Therefore, the invention provides a new treatment thought and method for clinical treatment of tumors, especially gastric cancer, provides a lead compound for development of an LSD1 degradation agent, and has important guiding significance.

Description

Technical Field

[0001] The invention belongs to the technical field of biomedicine, and in particular relates to a new medical application of Ailanthone. Background Art

[0002] Gastric cancer is one of the most common malignant tumors in the world. In China, the incidence and mortality of gastric cancer are both ranked third. Surgical resection is the most effective treatment for patients with early gastric cancer. However, due to the lack of specific symptoms in patients with early gastric cancer and the difficulty in attracting enough attention to early symptoms, nearly 70% of patients have already developed to the advanced or late stage when they seek medical treatment. At present, there is still a lack of effective treatment for advanced gastric cancer. Therefore, finding a specific therapeutic drug for advanced gastric cancer has become an urgent problem to be solved.

[0003] Compared with traditional tumor chemotherapy drugs, new anti-tumor drugs have the characteristics of good efficacy, high selectivity, and good patient compliance. Therefore, in recent years, new anti-tumor drugs have gradually become a new means in the field of tumor treatment. Currently, new anti-tumor drugs on the market include large molecule monoclonal antibody drugs and small molecule targeted drugs. Among them, large molecule monoclonal antibody drugs such as bevacizumab, trastuzumab, and nivolumab, and small molecule targeted drugs such as erlotinib and regorafenib for the treatment of digestive system tumors are already on the market. In addition to regorafenib, which is a multi-target tyrosine kinase inhibitor that can antagonize targets such as vascular endothelial growth factor receptor 1-3 (VEGFR-1, VEGFR-2, VEGFR-3), tyrosine kinase receptor (TIE-2), and serine / threonine protein kinase (V-Raf murine sarcomaviral oncogene homolog B1, BRAF), other drugs specifically act on targets such as vascular endothelial growth factor (VEGF), human epidermal growth factor receptor 2 (HER2), programmed death-1 (PD-1), and epidermal growth factor receptor (EGFR).

[0004] Compared with traditional chemotherapy drugs, new anti-tumor drugs have clear targets, act on tumor cells, reduce the killing effect on normal cells, and significantly reduce systemic toxic side effects. However, the current new anti-tumor drugs mainly focus on targets that are relatively well-known, such as the above-mentioned EGFR, HER-2, VEGF / VEFGR, BRAF, ALK, c-met, BCR-ABL and other genes that are overexpressed or easily mutated in tumor cells, as well as key regulatory molecules that regulate the tumor microenvironment and related immune systems, such as PD-1 / PD-L1, CD47, LAG3, etc. Therefore, there is a large gap in new anti-tumor drugs, and new targets and new mechanism drugs with high efficiency and low toxicity are urgently needed.

[0005] LSD1 is a flavin adenine dinucleotide (FAD)-dependent histone lysine-specific demethylase that can participate in a variety of cellular processes such as cell proliferation, differentiation, apoptosis, epithelial-mesenchymal transition, and angiogenesis by controlling the methylation status of histones or non-histones. LSD1 is highly expressed in a variety of tumor tissues such as prostate cancer, lung cancer, breast cancer, and gastric cancer, and promotes the proliferation, invasion, and metastasis of cancer cells by regulating the expression of tumor-related genes. Current studies have shown that the abnormal expression of LSD1 in hematological tumors and various solid tumors is closely related to the grade, malignancy, and prognosis of the tumor, suggesting that LSD1 is a potential therapeutic target for a variety of cancers.

[0006] Currently, several LSD1 inhibitors have entered clinical research. However, some LSD1 inhibitors still have high affinity for multiple other targets, often accompanied by serious adverse reactions. Some inhibitors have terminated clinical research due to large side effects or unsatisfactory efficacy. Compared with traditional small molecule inhibitors, degraders have significant advantages in affecting the non-enzymatic activity of target proteins, overcoming tumor resistance, and degrading undruggable targets. Therefore, it is of great significance to develop degraders for the histone demethylase LSD1, but currently reports on its degraders are limited to UM171 and BEA-17, and there are no reports of other targeted LSD1 degradation drugs.

[0007] Ailanthone is one of the main active ingredients extracted and separated from the dry stem bark or root bark of Ailanthus altissima, a plant of the Simaroubaceae family. Modern pharmacological experiments have shown that Ailanthone has antiviral and anti-tumor activities. However, there are few reports on Ailanthone degrading LSD1 protein, and there are no reports on Ailanthone as a LSD1 degrader to treat tumors. Summary of the invention

[0008] In view of this, an object of the present invention is to provide a use of Ailanthone in the preparation of an LSD1 protein degrading agent. Further, Ailanthone can significantly degrade LSD1 protein in tumor cells in a concentration-dependent and time-dependent manner. Wherein, the tumor cells are preferably gastric cancer cells, esophageal cancer cells, lung cancer cells, liver cancer cells or breast cancer cells.

[0009] The "LSD1 protein degrading agent" in the present invention mainly refers to drugs that degrade LSD1 protein, including Ailanthus ketone.

[0010] Among them, Ailanthone is an active ingredient in Ailanthus altissima, which can be extracted from the medicinal plant Ailanthus altissima. Its chemical structure is:

[0011] The second object of the present invention is to provide an application of a LSD1 protein degrading agent in the preparation of LSD1 targeted drugs.

[0012] The third object of the present invention is to provide a use of an LSD1 protein degrading agent in the preparation of a drug for preventing and / or treating a disease associated with abnormal LSD1 activity or expression. Wherein, the disease associated with abnormal LSD1 activity or expression is a tumor. The tumor is preferably gastric cancer, and specifically, the gastric cancer cell is preferably at least one of MKN-45 cells, AGS cells, MGC-803 cells and HGC-27 cells.

[0013] The fourth object of the present invention is to provide a use of a LSD1 protein degrading agent in the preparation of a product having any of the following functions:

[0014] 1) Degradation of LSD1 protein;

[0015] 2) Inhibit tumor cell proliferation and clone formation;

[0016] 3) Inhibit the growth of tumor cell xenografts.

[0017] Preferably, the LSD1 protein degrader is used in the preparation of a method for inhibiting the growth of nude mice bearing human gastric cancer MKN-45 cell transplanted tumors and nude mice bearing human gastric cancer MGC-803 cell transplanted tumors.

[0018] Therefore, Ailanthus ketone can significantly degrade LSD1 protein in a dose-dependent and time-dependent manner, and is an effective LSD1 protein degrader. It has a strong killing effect on tumor cells, especially gastric cancer cells, and has a strong inhibitory activity on tumor cell proliferation; Ailanthus ketone has a significant monoclonal formation inhibition ability on various tumor cell lines such as gastric cancer in vitro, and can inhibit the growth of various tumor cell transplants such as gastric cancer in vivo, and is expected to be used in the treatment of various tumors, especially gastric cancer. Therefore, the present invention provides a new treatment idea and method for clinical tumor treatment, and provides a lead compound for the development of LSD1 degraders, which has important guiding significance. BRIEF DESCRIPTION OF THE DRAWINGS

[0019] Figure 1 The graph shows the effect of different concentrations of Ailanthus on the proliferation of esophageal cancer KYSE-150 and TE-1 cells, gastric cancer HGC-27 and MGC-803 cells, lung cancer A549 and H1299 cells, liver cancer HepG2 cells and breast cancer MCF-7 cells after treatment for 72 hours.

[0020] Figure 2 The figure shows the result that Ailanthus altissima degrades LSD1 protein in gastric cancer MKN-45 cells in a concentration-dependent manner.

[0021] Figure 3 The figure shows the result that Ailanthus altissima degrades LSD1 protein in gastric cancer AGS cells in a concentration-dependent manner.

[0022] Figure 4 The figure shows the result that Ailanthus altissima degrades LSD1 protein in gastric cancer MGC-803 cells in a concentration-dependent manner.

[0023] Figure 5 The figure shows the result that Ailanthus altissima degrades LSD1 protein in gastric cancer HGC-27 cells in a concentration-dependent manner.

[0024] Figure 6 The figure shows the result that Ailanthus altissima degrades LSD1 protein in lung cancer H1299 cells in a concentration-dependent manner.

[0025] Figure 7 The figure shows the result that Ailanthone degrades LSD1 protein in lung cancer A549 cells in a concentration-dependent manner.

[0026] Figure 8 The figure shows the result that Ailanthone degrades LSD1 protein in esophageal cancer TE-1 cells in a concentration-dependent manner.

[0027] Fig. 9 The figure shows the result that Ailanthus altissima degrades LSD1 protein in liver cancer HepG2 cells in a concentration-dependent manner.

[0028] Fig.10The figure shows the result that Ailanthus altissima degrades LSD1 protein in breast cancer MCF-7 cells in a concentration-dependent manner.

[0029] Fig.11 The figure shows the result that Ailanthus altissima degrades LSD1 protein in gastric cancer MKN-45 cells in a time-dependent manner.

[0030] Fig.12 The figure shows the result that Ailanthus altissima degrades LSD1 protein in gastric cancer AGS cells in a time-dependent manner.

[0031] Fig.13 The graph shows the results of Ailanthus altissima inhibiting the proliferation of gastric cancer MKN-45, AGS, MGC-803 and HGC-27 cells in a dose-dependent and time-dependent manner.

[0032] Fig.14 The graph shows the results of the experiment on the inhibition of gastric cancer MKN-45, AGS, MGC-803 and HGC-27 cell clone formation by Ailanthus altissima. In the graph, * represents p<0.05, ** represents p<0.01, and *** represents p<0.001.

[0033] Fig.15 The graphs show the results of Ailanthus altissima inhibiting tumors in nude mice bearing human gastric cancer MKN-45 cell transplants and human gastric cancer MGC-803 cell transplants, where *** in the graph represents p<0.001. DETAILED DESCRIPTION

[0034] The technical solution of the present invention is further described in detail below through specific implementation methods.

[0035] Unless otherwise specified, the terms used in the present invention are commonly used in the field. The technical means used in the examples, such as preparation processes, test methods, etc., are all conventional means well known to those skilled in the art, and the reagents and products used are all commercially available. The sources and trade names of the reagents used, as well as the components thereof, are indicated when they first appear.

[0036] The tumor cells and Ailanthus ketone solutions used in the following examples are as follows:

[0037] Tumor cell line culture: human gastric cancer cell MKN-45 used RPMI-1640 culture medium containing 10% Gibco and 1% double antibody, esophageal cancer cell KYSE-150, esophageal cancer cell TE-1, human gastric cancer cell AGS, human gastric cancer cell HGC-27, lung cancer cell A549 and lung cancer H1299 used RPMI-1640 culture medium containing 10% FBS and 1% double antibody, human gastric cancer cell MGC-803, liver cancer cell HepG2 and breast cancer cell MCF-7 used RPMI-DMEM culture medium containing 10% FBS and 1% double antibody, and the above cells were cultured in a 37°C constant temperature incubator containing 5% CO2.

[0038] Preparation of Ailanthone solution: First, accurately weigh 1.00 mg of Ailanthone powder (purity: 99.76%) and place it in a 1.5 mL brown tube. Use a pipette to accurately measure 53 μL of DMSO (dimethyl sulfoxide) solution and add it to the tube. Vortex it thoroughly to completely dissolve it to prepare an Ailanthone solution with a final concentration of 50 mM. Store it in a -20°C refrigerator for later use.

[0039] Example 1. Ailanthus altissima inhibits cell viability of various tumor cells

[0040] Esophageal cancer cells KYSE-150 and TE-1, gastric cancer cells HGC-27 and MGC-803, lung cancer cells A549 and H1299, liver cancer cells HepG2 and breast cancer cells MCF-7 were plated in 96-well plates. After the cells adhered to the wall, Ailanthone diluted with culture medium to different concentrations was added to the 96-well plate. The concentration gradient of Ailanthone was: 0μM, 0.078μM, 0.156μM, 0.313μM, 0.625μM, 1.25μM, 2.5μM, 5μM, 10μM. After 72h of action, the above 96-well plate was taken out of the incubator and 20μL / well, 5mg / mL MTT (thiazolyl blue) solution was added in a light-proof clean bench, and then put into the incubator again for another 4h. After the incubation, the supernatant was discarded into the waste liquid bucket, 150 μL of DMSO solution was added to each well, and the well was placed on a shaker and shaken rapidly for 15 min. After the crystals were fully dissolved, the absorbance value was measured at a wavelength of 570 nm using an ELISA reader. Among them, cell inhibition rate (%) = (1-OD value of the drug group / OD value of the negative control group) × 100%, and the half inhibition rate IC was calculated using GraphPad Prism software 50 The results are shown in Table 1 and Figure 1 shown.

[0041] Table 1 Effects of Ailanthus altissima on cell viability of various tumor cells

[0042]

[0043] Figure 1 The results show the effects of different concentrations of Ailanthus ketone on the cell viability of various tumor cells after 72 hours of treatment. It can be found that with the increase of Ailanthus ketone dose, the inhibitory effect of Ailanthus ketone on the above tumor cells is stronger, with obvious dose dependence. Figure 1 It can be found that Ailanthus altissima has a stronger inhibitory effect on gastric cancer cell lines.

[0044] Example 2: Experiment on the degradation of LSD1 protein in different tumor cells by Ailanthus altissima

[0045] 2.1 Effects of different concentrations of Ailanthus angustifolia on LSD1 protein degradation in tumor cells

[0046] First, treat MKN-45, AGS, MGC-803, HGC-27, H1299, A549, TE-1, HepG2 and MCF-7 tumor cells with different concentrations of Ailanthus ketone for 24 hours, collect the cell extract protein, and quantify the protein concentration using a BCA kit; then take an equal amount of protein and use 10% separation gel for protein immunoblotting. After the electrophoresis, transfer the protein to the NC membrane, and block it with 5% skimmed milk powder for 2 hours after the transfer. After blocking, incubate the target bands with primary antibodies of LSD1 and GAPDH (glyceraldehyde-3-phosphate dehydrogenase) at 4°C overnight. On the next day, wash the bands with TBST and incubate them with secondary antibodies of LSD1 and GAPDH at room temperature for 2 hours. After the secondary antibody incubation, wash the bands with TBST, then evenly cover the luminescent liquid on the bands, and use a WB exposure instrument for imaging. The results are as follows: Figures 2 to 10 shown.

[0047] Among them, the specific samples of protein immunoblotting experiment are as follows:

[0048] MKN-45 cells were treated with Ailanthone at concentrations of 0.1 μM, 0.2 μM, 0.4 μM, 0.8 μM, 1.6 μM, 3.2 μM, and 6.4 μM;

[0049] AGS and TE-1 cells were treated with Ailanthone at concentrations of 0.05 μM, 0.1 μM, 0.2 μM, 0.4 μM, 0.8 μM, 1.6 μM, and 3.2 μM, respectively;

[0050] MGC-803 and HGC-27 cells were treated with Ailanthone at concentrations of 0.06 μM, 0.1 μM, 0.25 μM, 0.5 μM, 1 μM, 2 μM, and 4 μM, respectively;

[0051] H1299 and MCF-7 cells were treated with Ailanthone at concentrations of 0.15 μM, 0.3 μM, 0.6 μM, 1.2 μM, 2.4 μM, 4.8 μM, and 9.6 μM, respectively;

[0052] A549 cells were treated with Ailanthone at concentrations of 0.2 μM, 0.4 μM, 0.8 μM, 1.6 μM, 3.2 μM, 6.4 μM, and 12.8 μM;

[0053] HepG2 cells were treated with Ailanthone at concentrations of 0.25 μM, 0.5 μM, 1 μM, 2 μM, 4 μM, 8 μM and 16 μM, respectively.

[0054] Figure 2 This indicates that Ailanthone can reduce the level of LSD1 protein in gastric cancer MKN-45 cells in a dose-dependent manner. Combined with the fitting curve of Ailanthone concentration and LSD1 degradation rate shown in the figure, the DC of Ailanthone on LSD1 protein in gastric cancer MKN-45 cells was calculated. 50 (drug concentration at which 50% of the target protein is degraded) is 1.24 μM.

[0055] Figure 3 This indicates that Ailanthus ketone can reduce the level of LSD1 protein in gastric cancer AGS cells in a dose-dependent manner. Combined with the fitting curve of Ailanthus ketone concentration and LSD1 degradation rate shown in the figure, the DC of Ailanthus ketone on LSD1 protein in gastric cancer AGS cells was calculated. 50 It is 0.28μM.

[0056] Depend on Figure 4 It can be seen that with the increase of Ailanthone dosage, the degradation effect of Ailanthone on LSD1 protein in gastric cancer MGC-803 cells became stronger. Combining the fitting curve of Ailanthone concentration and LSD1 degradation rate shown in the figure, the DC of Ailanthone on LSD1 protein in gastric cancer MGC-803 cells was calculated. 50 It is 0.36μM.

[0057] Figure 5 The results showed that Ailanthone could reduce the level of LSD1 protein in gastric cancer HGC-27 cells in a dose-dependent manner. Combined with the fitting curve of Ailanthone concentration and LSD1 degradation rate shown in the figure, the DC of Ailanthone on LSD1 protein in gastric cancer HGC-27 cells was calculated. 50 It is 0.37μM.

[0058] Figure 6 This indicates that Ailanthone can reduce the level of LSD1 protein in lung cancer H1299 cells in a dose-dependent manner. Combined with the fitting curve of Ailanthone concentration and LSD1 degradation rate shown in the figure, the DC of Ailanthone on LSD1 protein in lung cancer H1299 cells was calculated. 50 It is 0.27μM.

[0059] Figure 7This indicates that Ailanthone can reduce the level of LSD1 protein in lung cancer A549 cells in a dose-dependent manner. Combined with the fitting curve of Ailanthone concentration and LSD1 degradation rate shown in the figure, the DC of Ailanthone on LSD1 protein in lung cancer A549 cells was calculated. 50 It is 0.38μM.

[0060] Depend on Figure 8 It can be seen that with the increase of Ailanthone dosage, the degradation effect of Ailanthone on LSD1 protein in esophageal cancer TE-1 cells became stronger. Combining the fitting curve of Ailanthone concentration and LSD1 degradation rate shown in the figure, the DC of Ailanthone on LSD1 protein in esophageal cancer TE-1 cells was calculated. 50 It is 0.59μM.

[0061] Fig. 9 This indicates that Ailanthone can reduce the level of LSD1 protein in HepG2 liver cancer cells in a dose-dependent manner. Combined with the fitting curve of Ailanthone concentration and LSD1 degradation rate shown in the figure, the DC of Ailanthone on LSD1 protein in HepG2 liver cancer cells was calculated. 50 0.11μM.

[0062] Fig.10 This indicates that Ailanthone can reduce the level of LSD1 protein in breast cancer MCF-7 cells in a dose-dependent manner. Combined with the fitting curve of Ailanthone concentration and LSD1 degradation rate shown in the figure, the DC of Ailanthone on LSD1 protein in breast cancer MCF-7 cells was calculated. 50 It is 2.04μM.

[0063] 2.2 Effects of Ailanthus ketone treatment for different time periods on LSD1 protein degradation in gastric cancer cells

[0064] First, gastric cancer cells were treated with Ailanthus altissima for different periods of time, and then the cells were collected to extract total protein, and the protein concentration was quantified using a BCA kit; then an equal amount of protein was taken for protein immunoblotting using a 10% separation gel. After the electrophoresis, the protein was transferred to an NC membrane, and after the transfer, it was blocked with 5% skimmed milk powder for 2 hours. After blocking, the target bands were incubated with the primary antibodies of LSD1 and GAPDH at 4°C overnight. On the next day, the bands were washed with TBST and incubated with the secondary antibodies of LSD1 and GAPDH at room temperature for 2 hours. After the secondary antibody incubation, the bands were washed with TBST, and then the luminescent liquid was evenly covered on the bands, and the WB exposure instrument was used for imaging. The results are shown as follows: Figures 11-12 shown.

[0065] Among them, the specific samples of protein immunoblotting experiment are as follows:

[0066] MKN-45 cells were treated with 0.8 μM Ailanthone for 2 h, 6 h, 12 h, 24 h, 36 h, and 48 h;

[0067] AGS cells were treated with 0.4 μM Ailanthone for 2 h, 6 h, 12 h, 24 h, 36 h and 48 h, respectively.

[0068] Fig.11 This shows that the longer Ailanthone acts on gastric cancer MKN-45 cells, the stronger its inhibitory effect on LSD1 protein; Fig.12 This shows that Ailanthus altissima could reduce the LSD1 protein level in gastric cancer AGS cells in a time-dependent manner.

[0069] therefore, Figures 2 to 12 The results showed that Ailanthus altissima could degrade LSD1 protein in tumors in a dose-dependent and time-dependent manner, and Ailanthus altissima could be used as an effective degrader of LSD1.

[0070] Example 3: Ability of Ailanthus ketone to inhibit proliferation of gastric cancer cells

[0071] MKN-45 cells, AGS cells, MGC-803 cells and HGC-27 cells were inoculated into three 96-well plates at 4500 cells / well, 4000 cells / well and 3500 cells / well, respectively. After the cells adhered to the wall, different concentrations of Ailanthone diluted with culture medium were added to the 96-well plates. The concentration gradient of Ailanthone was: 0μM, 0.15625μM, 0.3125μM, 0.625μM, 1.25μM, 2.5μM, 5μM, 10μM, 20μM. After culturing for 24h, 48h and 72h, the 96-well plates were taken out, 20μL / well, 5mg / mL MTT solution was added in a light-proof clean bench, and then the plates were placed in the incubator again for 4h. After the incubation, the supernatant was discarded into the waste liquid bucket, 150 μL of DMSO solution was added to each well, and the well was placed on a shaker and shaken rapidly for 15 minutes. After the crystals were fully dissolved, the absorbance value was measured at a wavelength of 570 nm using an ELISA reader. Cell inhibition rate (%) = (1-OD value of the drug group / OD value of the negative control group) × 100%, and the half inhibition rate IC was calculated using GraphPad Prism software 50 , the results are as follows Fig.13 shown.

[0072] from Fig.13 It was found that Ailanthus ketone can significantly inhibit the cell viability of gastric cancer cells MKN-45, AGS, MGC-803 and HGC-27, and Ailanthus ketone has a strong killing effect on gastric cancer cells when acting for 24h, 48h and 72h respectively; and the higher the concentration of Ailanthus ketone and the longer the action time, the stronger the ability to inhibit the proliferation of gastric cancer cells. Therefore, Ailanthus ketone has the ability to inhibit the proliferation of gastric cancer cells as a LSD1 degrader.

[0073] Example 4: Ailanthus altissima inhibits the colony-forming ability of gastric cancer cells

[0074] MKN-45 cells, AGS cells, MGC-803 cells, and HGC-27 cells were inoculated in 6-well plates at 1000 cells / well. After the cells adhered to the wall, the culture medium containing different concentrations of Ailanthone was replaced and cultured for 24 hours. After washing with PBS twice, the culture medium was replaced with fresh culture medium and placed in an incubator for 2 weeks. The culture medium was changed every two days. The culture was terminated when obvious colonies were formed. Finally, the cells were fixed with 4% paraformaldehyde, stained with crystal violet, and quantified with Image J software. GraphPad Prism software was used for statistical analysis. The results are shown in Figure 2. Fig.14 shown.

[0075] Fig.14 The results showed that, compared with the control group, different concentrations of Ailanthone treatment could effectively inhibit the cloning ability of gastric cancer cells MKN-45, AGS, MGC-803 and HGC-27. Moreover, as the dose of Ailanthone increased, the inhibitory effect on the formation of gastric cancer cell clones became stronger. Therefore, Ailanthone, as a LSD1 degrader, also has the ability to inhibit the cloning of gastric cancer cells.

[0076] Example 5: Study on the anti-gastric cancer activity of Ailanthus ketone in vivo

[0077] The nude mice were kept in an SPF environment for 3-5 days. The gastric cancer cells MKN-45 and MGC-803 were cultured and collected, and the cells were suspended in saline. 200 μL of the cell suspension was inoculated under the armpit of each nude mouse. Each nude mouse was inoculated with 6×10 6 After inoculation of cells, the growth of the transplanted tumor was observed. When the volume of the transplanted tumor reached about 80 mm 3 At about 14:00 p.m., the nude mice were divided into groups according to the tumor volume, with six mice in each group. After the grouping was completed, intraperitoneal administration began, and the MKN-45 tumor-bearing mice groups were set as the solvent control group and the 5.0 mg·kg -1 ·day -1 Drug administration group: MGC-803 tumor-bearing mice were divided into solvent control group and 4.0 mg·kg -1 ·day -1 The drug group was compared with 8.0 mg·kg -1 ·day -1 The drug group was given once every 2 days for 2 weeks. When the maximum volume of the transplanted tumor in the solvent control group reached 1200mm 3 The animal experiment was terminated at about 1:10 pm. Fig.15 shown.

[0078] from Fig.15It can be found that Ailanthus ketone has a significant inhibitory effect on the growth of nude mice bearing human gastric cancer MKN-45 cell transplants and nude mice bearing MGC-803 cell transplants. In the nude mouse bearing MGC-803 cell transplant model, it was found that Ailanthus ketone's inhibitory effect on transplanted tumors was dose-dependent, further demonstrating Ailanthus ketone's good in vivo anti-tumor effect. Therefore, Ailanthus ketone, as an LSD1 degrader, also has a good anti-tumor effect in vivo.

[0079] Therefore, Ailanthus altissima can significantly degrade LSD1 protein in gastric cancer cells MKN-45 and AGS in a dose-dependent and time-dependent manner, and has a strong killing effect on gastric cancer cells, has a strong inhibitory activity on gastric cancer cell proliferation, can inhibit the growth of gastric cancer cell xenografts, and is expected to be used in the treatment of gastric cancer.

[0080] In summary, the present invention provides a new treatment idea and method for the clinical treatment of gastric cancer and other tumors, and has important guiding significance for the development of LSD1 degraders.

[0081] Finally, it should be noted that the above embodiments are only used to illustrate the technical solution of the present invention rather than to limit it. Although the present invention has been described in detail with reference to the preferred embodiments, ordinary technicians in the field should understand that the specific implementation methods of the present invention can still be modified or some technical features can be replaced by equivalents without departing from the spirit of the technical solution of the present invention, which should be included in the scope of the technical solution for protection of the present invention.

Claims

1. An application of Ailanthus ketone in the preparation of an LSD1 protein degradation agent, wherein: The chemical structure of Ailanthus ketone is:

2. The use according to claim 1, characterized in that: Ailanthone degraded LSD1 protein in tumor cells in a concentration-dependent and time-dependent manner; Preferably, the tumor cells are gastric cancer cells, esophageal cancer cells, lung cancer cells, liver cancer cells or breast cancer cells.

3. Use of a LSD1 protein degrader in the preparation of a LSD1 targeted drug, wherein: The LSD1 protein degradation agent includes Ailanthus ketone.

4. The use according to claim 3, characterized in that: Application of Ailanthus altissima in the preparation of a drug for preventing and / or treating diseases associated with abnormal LSD1 activity or expression.

5. The use according to claim 4, characterized in that: The disease associated with abnormal LSD1 activity or expression is a tumor.

6. The use according to claim 5, characterized in that: The tumor is gastric cancer, esophageal cancer, lung cancer, liver cancer or breast cancer.

7. The use according to claim 6, characterized in that: The tumor cells are esophageal cancer cells KYSE-150, esophageal cancer cells TE-1, gastric cancer cells HGC-27, gastric cancer cells MGC-803, gastric cancer cells MKN-45, gastric cancer cells AGS, lung cancer cells A549, lung cancer cells H1299, liver cancer cells HepG2 or breast cancer cells MCF-7.

8. Use of a LSD1 protein degradation agent in the preparation of a product having any of the following functions, wherein: The LSD1 protein degradation agent includes Ailanthus ketone: 1) Degradation of LSD1 protein; 2) Inhibit tumor cell proliferation and clone formation; 3) Inhibit the growth of tumor cell xenografts.

9. The use according to claim 8, characterized in that: The invention discloses a method for preparing a LSD1 protein degrading agent for inhibiting the growth of nude mouse-bearing human gastric cancer MKN-45 cell transplanted tumors and nude mouse-bearing human gastric cancer MGC-803 cell transplanted tumors.

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