Small-molecule compound, non-covalent CRM1 inhibitor and application of non-covalent CRM1 inhibitor

Abstract

The invention discloses a small molecule compound, a non-covalent CRM1 inhibitor and application of the non-covalent CRM1 inhibitor, and belongs to the technical field of medicine. According to the invention, compounds 1-9 of non-covalent binding CRM1, which have stronger affinity, are optimally designed and synthesized on the basis of earlier-stage research results, and are subjected to an in-vitro pull-down experiment, an immunofluorescence experiment and a cell proliferation experiment; it is found that the compound 3 and the compound 8 show in-vitro inhibition of formation of CRM1 nucleation compounds, in-vivo obvious inhibition of CRM1 mediated cargo nucleation and inhibition of growth of Hela cells and A549 cells. Therefore, the small molecule compound has a good inhibition effect on CRM1, can be used for inhibiting growth of various cancer cells, and has relatively high potential clinical application value and development prospect.

Description

Technical Field

[0001] The present invention relates to the field of medical technology, and in particular to a small molecule compound, a non-covalent CRM1 inhibitor and applications thereof. Background Art

[0002] Although cancer treatments have greatly advanced, effective therapies remain lacking for some tumor subtypes, as well as for the vast majority of drug-resistant and advanced tumors. Given the increasing incidence of cancer, the high mortality risk, and the increasing cost of treatment, the development of low-toxic, highly effective, and broad-spectrum anti-cancer drugs would be of great benefit. Recent studies have demonstrated that specific inhibitors of the nuclear export factor CRM1 (Chromosome Region Maintenance 1, also known as exportin 1 or XPO1) exhibit potent and broad-spectrum anti-tumor activity. CRM1 is an essential nuclear export factor in the human body, whose primary function is to transport proteins containing the nuclear export signal (NES) from the nucleus to the cytoplasm through the nuclear pore. It plays a crucial role in the cell cycle, particularly in regulating cell growth, and apoptosis. CRM1 plays a crucial role in tumor development and progression, particularly in regulating the nuclear transport of tumor suppressor proteins. CRM1 inhibitors are being developed as a novel therapeutic strategy, potentially offering new treatment options for cancer patients.

[0003] Numerous studies have been conducted on CRM1 inhibitors. The first-generation CRM1 inhibitor, LMB, was discovered to covalently bind to CRM1, relying on the conserved Cys528 of CRM1 to exert its therapeutic effects. Its binding to CRM1 is irreversible, resulting in high toxicity and has been discontinued. Second-generation CRM1 inhibitors, SINE compounds, are similar to first-generation CRM1 inhibitors, still covalently binding to CRM1 but with increased reversibility, resulting in lower toxicity. For example, KPT-330 received US FDA approval in 2019 for the treatment of relapsed or refractory multiple myeloma. However, despite these improvements, the toxic side effects of these covalent CRM1 inhibitors remain significant, primarily due to strong gastrointestinal reactions. Therefore, the development of a non-covalent CRM1 inhibitor with high stability, enhanced efficacy, and low toxicity is highly desirable. Summary of the Invention

[0004] In order to address the above-mentioned deficiencies in the prior art, the present invention aims to provide a structure-based broad-spectrum anti-tumor non-covalent CRM1 inhibitor and its application, so as to obtain a non-covalent CRM1 inhibitor with high stability, strong efficacy and low toxicity.

[0005] The technical solution of the present invention to solve the above technical problems is as follows: a small molecule compound is provided, and the molecular structure of the small molecule compound is shown in formula (I) or formula (II):

[0006] (I)

[0007] (II).

[0008] The present invention provides a use of the above-mentioned small molecule compound in the preparation of a non-covalent CRM1 inhibitor and / or a drug for preventing and / or treating a disease associated with CRM1.

[0009] Furthermore, diseases associated with CRM1 include pancreatic cancer, endometrial cancer, colorectal cancer, breast cancer, ovarian cancer, lung cancer, and cervical cancer.

[0010] The present invention provides a non-covalent CRM1 inhibitor, which includes at least one of the above-mentioned small molecule compounds.

[0011] The present invention also provides a broad-spectrum anti-tumor drug, which is a preparation prepared with the above-mentioned small molecule compound or the above-mentioned non-covalent CRM1 inhibitor as an active ingredient and pharmaceutically acceptable excipients or auxiliary ingredients.

[0012] Furthermore, the broad-spectrum anti-tumor drug can be administered orally, by injection, by rectal perfusion, by sublingual administration, by inhalation, by topical administration in vitro, or by systemic transdermal administration.

[0013] Furthermore, the dosage forms of broad-spectrum anti-tumor drugs include injections, capsules, tablets, sustained-release preparations, suppositories and sprays.

[0014] The present invention has the following beneficial effects: (1) In order to obtain small molecules with better inhibitory effects on CRM1, the present invention further modified the compounds based on the results of previous experiments: adding hydrogen bonds or hydrophobic interactions to the compounds, optimizing the shape fit of the binding interface, reducing steric hindrance, and simplifying groups that have no effect on the interaction. Compounds 1-9 were designed and synthesized, thereby improving the affinity of the compounds to CRM1.

[0015] (2) Through in vitro pull-down assays, immunofluorescence assays, and cell proliferation assays, the present invention found that compounds 3 and 8 inhibited the formation of CRM1 nuclear export complexes in vitro, significantly inhibited CRM1-mediated cargo nuclear export in vivo, and were able to inhibit the growth of Hela and A549 cells. Therefore, the small molecule compounds of the present invention have a good inhibitory effect on CRM1 and can be used to inhibit the growth of various cancer cells, with high potential clinical application value and development prospects. BRIEF DESCRIPTION OF THE DRAWINGS

[0016] Figure 1 is the molecular structural formula of compound 1-9; Figure 2 The UV, total ion and mass spectra of compound 3; Figure 3 is the NMR spectrum of compound 3; Figure 4 The UV, total ion and mass spectra of compound 8; Figure 5 Graph showing the in vitro inhibitory effects of compounds 1-9 on CRM1; Figure 6 The graph shows the inhibitory effect of 100 μM compounds 1-9 on CRM1-mediated nuclear export; Figure 7 The graph shows the inhibitory effect of 50 μM compounds 1-9 on CRM1-mediated nuclear export; Figure 8 The graph shows the inhibitory effect of 20 μM compound 2-8 on CRM1-mediated nuclear export; Figure 9 Graph showing the growth inhibition effect of compound 1-9 on Hela cells; Figure 10 Graph showing the growth inhibitory effect of compounds 1-9 on A549 cells. DETAILED DESCRIPTION

[0017] The following examples are intended only to illustrate the present invention and are not intended to limit the scope of the present invention. In the examples, where specific conditions are not specified, the experiments were performed under conventional conditions or those recommended by the manufacturer. Reagents or instruments used, where the manufacturer is not specified, are commercially available conventional products.

[0018] Experimental methods: (1) Cell culture The present invention mainly uses Hela cells, and the cell culture medium is DMEM medium containing 10wt% FBS and 1wt% double antibiotic (penicillin-streptomycin), which is cultured in a 37°C cell culture incubator containing 5wt% CO2, and is usually passaged every 3 days.

[0019] ① Cell recovery: Open the 37°C water bath in advance, take out the cells from the liquid nitrogen tank or -80°C freezer, and gently shake them in the water bath to thaw them quickly; then transfer the thawed cells from the cryopreservation tube to a 5mL EP tube, quickly add 4mL of fresh culture medium to dilute the cryopreservation solution, balance and centrifuge; discard the supernatant of the centrifuged cells, add 1mL of culture medium to resuspend the cells, gently blow off the cells, and transfer them to a 10cm cell culture dish filled with 10mL of culture medium in advance. Finally, place the cell culture dish in a cell incubator for culture; ②Cell passaging: Spray the sterile 1×PBS solution, cell culture medium, 25wt% trypsin, and a new cell culture dish with 75wt% alcohol required for the passaging process and place it in a biosafety cabinet for UV irradiation for 30 minutes; then take out the cells, discard the old culture medium, add 2mL of 1×PBS, gently inject along the wall to avoid impacting the cells, shake gently to wash away the residual culture medium on the cell surface, and discard the PBS; then add 2mL of 25wt% trypsin, shake gently to evenly distribute it on the cell surface, and wait for the cells to digest; during the waiting process, 10mL of new culture medium can be added to the new cell culture dish in advance, and you can judge whether the digestion is complete by gently blowing on the cells to see if small holes appear; after the cells are completely digested, discard the trypsin, add 3mL of new culture medium, gently blow on the cells to resuspend the cells, and then take a certain proportion of cells and place them in a new culture dish for further culture; ③Cell freezing: After trypsinizing the cells according to the cell passaging method, add 1× PBS to resuspend the cells, centrifuge, discard the supernatant, and then add cell freezing solution. Gently blow the cells apart, and then transfer the cells to sterile cell freezing tubes, 1 mL per tube, and mark them (cell type and date). First, freeze them in a -80℃ freezer overnight, and then place them in a liquid nitrogen tank the next day. All the above operations should be performed in a biological safety cabinet and strictly follow sterile procedures; ④Cell transfection: Before transfection, cells need to be plated one day in advance, that is, subcultured to the required transfection well plate, so that the cell density reaches an appropriate level and is in the logarithmic phase of growth. Prepare two EP tubes and anti-serum-free cell culture medium and place them in a biosafety cabinet for UV irradiation. Then take out the plasmid and melt it in advance, and calculate the amount of plasmid and transfection reagent to be added. Generally speaking, the amount of transfection reagent added is twice that of the exogenous plasmid. Add plasmid to one EP tube and transfection reagent to the other EP tube, and then add the corresponding amount of anti-serum-free culture medium to each of them, mix gently, and let it stand for 3 minutes. After standing, gently mix the plasmid and transfection reagent and let it stand for 15 minutes. Take out the cell well plate that has been plated in advance, and evenly add the prepared plasmid and transfection reagent mixture to each well. For transfection of conventional overexpression plasmids, relevant experiments can be carried out after 24 hours.

[0020] (2) In vitro pull-down assay In vitro pull-down experiments can verify the interaction between proteins. The present invention uses pull-down experiments to verify the inhibitory effect of small molecule compounds on CRM1. The specific operation process is as follows: ① Remove the GST beads and equilibrate them in advance with GST Pull-down buffer (20mM Tris (pH 8.0), 150mM NaCl, 2mM MgCl2, 0.001wt% Triton X-100, 10wt% glycerol, 2mM dithiothreitol) three times; ② A certain amount of GST-NES was combined with GST beads at 4°C for 1.5 hours under rotation. At the same time, a certain amount of small molecule compound was incubated with CRM1 and Ran Q69L / L182A at room temperature for 1 hour. ③After GST beads bind to GST-NES, wash away the loosely bound GST NES with GST Pull-down buffer three times, then adjust the volume to the experimental needs and evenly add GST beads to the incubated protein and small molecule tubes; ④Place the sample in a 4℃ rotary pull-down chamber for 1.5h; ⑤ Take out the pull-down sample and wash it with GST pull-down buffer three times, then quantify it to the appropriate volume and add 10 μL loading sample; ⑥Put the sample in a metal bath and boil for 10 minutes. After centrifugation, load the sample and run SDS-PAGE gel electrophoresis to observe the phenomenon.

[0021] (3) Immunofluorescence experiment Cells were transfected with a plasmid expressing a fluorescently tagged protein, or the corresponding protein was incubated in cells with a fluorescent dye-labeled antibody. The subcellular localization and expression level of the protein were detected using a confocal microscope (transfection reagent: jetPRIME, purchased from Polyplus Transfection). The cells used in the immunofluorescence experiments of this invention were mainly Hela cells, and the specific procedures were as follows: ① Inoculate Hela cells into a 24-well plate pre-loaded with cell culture slides and transfect with the desired plasmid 24 hours later; ② 24 hours after transfection, add the drug at a pre-calculated concentration to the cells and prepare the drug solution using complete culture medium; ③ Fix the cells with pre-chilled methanol at -20°C for 15 min, then wash with pre-chilled PBS three times, 3 min each time; ④ Block with IF blocking solution (5wt% BSA) at room temperature for 30 minutes, then add 300 μL of diluted DAPI solution to each well and incubate for 7-10 minutes. Then, wash the slides three times with 1× PBS, each time for 3 minutes. ⑤ Seal the slides with anti-fluorescence quenching sealing agent and photograph them using a confocal scanning microscope.

[0022] (4) MTT assay for cell proliferation The MTT assay is a commonly used method for measuring cell proliferation activity in vitro. The specific procedure is as follows: ① Cell plating: MTT experiments are usually performed using 96-well plates. After cell digestion, 2,000 cells are added to each well by cell counting. The plate is then placed in a cell culture incubator and allowed to adhere. ② Adding drugs: Add the small molecule compound solution (compounds 1-9) to the EP tube, and then add cell culture medium (DMEM medium: 450mL; fetal bovine serum: 50mL; double antibody (penicillin and streptomycin): 5mL; mix thoroughly and store at 4℃) to make the final concentration of the small molecule compound solution 100µM and 20µM. Then add this solution to the 96-well plate of cells that have been plated in advance; ③Continue to culture: Place the 96-well plate with the drug added back into the cell culture incubator and continue to culture for 72 hours; ④ Fix the living cells; add 20µL of 0.5wt% MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) solution to each well using a syringe and return the well to the incubator for 1-4 hours. ⑤ Dissolve: Remove the fixed cells, discard the culture medium, and shake clean. Add 100µL of dimethyl sulfoxide to each well to dissolve the cells and place on a shaker to dissolve for 30 minutes. ⑥Absorbance detection: Measure the absorbance of cells at 570 nm using an enzyme-labeled instrument.

[0023] Example 1: Synthesis of Compound 3 and Compound 8 In order to obtain a small molecule with better inhibitory effect on CRM1, the present invention further modified the compound based on the co-crystal structure of CRM1 with the hope of improving the affinity of the compound with CRM1. The present invention added hydrogen bonds or hydrophobic interactions to the compound, optimized the shape fit of the binding interface, reduced steric hindrance, and simplified groups that had no effect on the interaction. Compounds 1-9 (chemical structures see Figure 1 ), the synthesis method thereof is as follows (only the synthesis process of compound 3 and compound 8 is given here): (1) The synthesis process of compound 3 is as follows:

[0024] ① Add tert-butyl 4-hydroxypiperidine-1-carboxylate (555 mg, 2.76 mmol), triphenylphosphine (723 mg, 2.76 mmol), and diisopropyl azodicarboxylate (558 mg, 2.76 mmol) to toluene (10 mL) and stir at 60°C under argon for 2 h. After the reaction, extract twice with ethyl acetate (50 mL), and the organic layer is dried and concentrated. Finally, the purified product is purified by silica gel column chromatography (EA / PE = 0-20%) as a white solid (390 mg, 70% yield); LC-MS (ESI) m / z: 407.0 [M+H]+; ② To the product obtained in step ① (390 mg, 0.96 mmol, dissolved in 10 mL of dioxane), add HCl / 1,4-dioxane (4 M, 10 mL). Stir at room temperature for 1 hour to obtain a crude product. Dichloromethane solution (10 mL) and NaOH / H₂O (5 M, 10 mL) were added to the crude product and stirred at room temperature for 0.5 hour until the reaction was complete. The suspension was then diluted with water (100 mL) and extracted twice with dichloromethane solution (50 mL). The organic layer was concentrated to obtain the reaction product as a white solid (230 mg, yield: 78%); LC-MS (ESI) m / z: 307.1 [M+H]⁺. ③ To the product obtained in step ② (230 mg, 0.75 mmol) and 3-fluoro-4-(trifluoromethyl)benzonitrile (425 mg, 2.25 mmol dissolved in 6 mL of dimethyl sulfoxide) was added N,N-diisopropylethylamine (290 mg, 2.25 mmol). The mixture was then stirred at 100°C for 36 hours. After completion of the reaction, the suspension was diluted with water (100 mL) and extracted twice with ethyl acetate (100 mL). The organic layer was dried and concentrated. The purified product was obtained by silica gel column chromatography (EA / PE = 0-25%) as a white solid (127 mg, 36% yield); LC-MS (ESI) m / z: 476.0 [M+H]+; ④ To the product from step ③ (127 mg, 0.27 mmol dissolved in 3 mL of ethanol), KOH / H₂O (5 M, 10 mL) was added, followed by stirring at 100°C for 5 hours. After completion of the reaction, the suspension was diluted with water (100 mL) and extracted twice with dichloromethane (50 mL). The organic layer was dried and concentrated. High-performance liquid chromatography (HPLC) [MeCN / H₂O (10 mmol / L HCOOH), X-Select 10 µm 19*250 mm, 20 mL / min, UV 254] was used to obtain compound 3 as a white solid (50.93 mg, 38% yield); LC-MS (ESI) m / z: 495 [M+H]⁺ (detection results see Table 1). Figure 2-3 ).

[0025] 1 H NMR (400 MHz, DMSO- d 6)δ 13.43(br,1H),8.93(dd, J =2.4Hz,1H),8.30(dd, J =6.8Hz,1H),8.04(s,1H),7.88–7.82(m,3H),7.60(q, J =4.0Hz,1H),7.49(d, J =2.0Hz,1H),4.97–4.93(m,1H),3.15–3.10(m,2H),3.00(t, J =8.8Hz,2H),2.17–2.14(m,2H),1.95–1.88(m,2H).

[0026] (2) The synthesis process of compound 8 is as follows:

[0027] ① To 3-fluoro-4-(trifluoromethyl)benzonitrile (2.0 g, 10.6 mmol) and 4-methylpiperidin-4-ol (dissolved in 20 mL of dimethyl sulfoxide, 1.46 g, 12.7 mmol) was added N,N-diisopropylethylamine (2.73 g, 21.2 mmol), and the mixture was stirred at 100°C for 36 h under nitrogen. Then, glacial NH4Cl solution (1 M 20 mL) was added and extracted twice with ethyl acetate. The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo to obtain the crude product, which was purified by flash column chromatography (PE / EA = 1 / 0~7 / 3, Silica-CS40g, 30 mL / min, silica gel, UV 214) to give the product as a yellow oil (2.1 g, yield 98%); ESI-MS m / z calculated for [C 14 H 15 F3N2O] [M+H]+:285.1;found:285.2; ② To the product obtained in step ① (dissolved in 100 mL of water, 6.1 g, 15.91 mmol) was added concentrated hydrochloric acid (100 mL), stirred at 100°C for 48 h, cooled to room temperature, and concentrated to dryness. The residue was purified by flash column chromatography (DCM / MeOH = 1 / 0 to 85 / 15, Silica-CS 40 g, 30 mL / min, silica gel, UV 214) to give a crude product, which was purified by preparative HPLC [MeCN / H2O (10 mmol / L HCOOH), X-Select 10µm 19*250mm, 20 mL / min, UV 254] to give a yellow solid (700 mg, yield 44%); ESI-MS m / z calcd for [C 14 H 16 F3NO3] [M+H] + :304.1;found:303.9; 1 H NMR (400 MHz, DMSO- d 6)δ 13.46(br,1H),8.00(s,1H),7.80(dd, J =17.2,8.4 Hz,2H),4.34(s,1H),3.07–3.01(m,2H),2.75–2.73(m,2H),1.61–1.58(m,4H),1.19(s,3H); ③ To the product from step ② (650 mg, 2.14 mmol dissolved in 10 mL of methanol) was added H₂SO₄ (3 drops) at 20°C. The mixture was then stirred at 70°C for 16 h. LCMS indicated consumption of the starting material and the desired mass spectrum was detected. The reaction mixture was concentrated in vacuo to yield a residue. Purification by flash column chromatography (PE / EA = 1 / 0 to 7 / 3, Silica-CS 20 g, 30 mL / min, silica gel, UV 214) afforded the product as a yellow oil (460 mg, 68% yield); ESI-MS m / z calculated for [C 15 H 18 F3NO3] [M+H]+:318.1;found:317.9; ④ To the product from step ③ (330 mg, 1.04 mmol dissolved in 10 mL of dichloromethane), Rh(OAc)2 (92 mg, 0.21 mmol) was added. Tert-butyl diazoacetate (296 mg, 2.08 mmol) was then added dropwise under nitrogen at 20°C. The mixture was stirred at 20°C for 16 h. LCMS showed approximately 60% of the starting material remained, and the desired compound was detected by mass spectrometry at approximately 30%. The reaction mixture was added to glacial NH4Cl (1 M 20 mL). The aqueous phase was extracted three times with dichloromethane (15 mL). The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo to obtain a residue. The residue was purified by flash column chromatography (PE / EA = 1 / 0 to 7 / 3, Silica-CS 12 g, 30 mL / min, silica gel, UV 214) to afford a yellow oil (153 mg, 34% yield). ESI-MS m / z calculated for [C 21 H 28 F3NO5] [M+H]+:432.2; found: 432.3. ⑤ To the product obtained in step ④ (150 mg, 0.35 mmol, dissolved in 5 mL of dichloromethane) was added trifluoroacetic anhydride (1.85 mL) at 0°C. The mixture was stirred at 20°C for 3 h and concentrated in vacuo to obtain a residue. The residue was purified by flash column chromatography (DCM / MeOH = 1 / 0 to 9 / 1, 12 g of Silica-CS, 30 mL / min, silica gel, UV 214) to afford compound 8 as a yellow oil (120 mg, 92% yield); ESI-MS m / z calculated for [C 17 H 20 F3NO5] [M+H]+:376.1; found:376.1 (test results see Figure 4 ).

[0028] Example 2: Inhibitory effect of compounds 1-9 on the formation of nuclear export complex by CRM1 For the compounds synthesized after structural optimization, their inhibitory effects on the formation of CRM1 nuclear export complex in vitro were first investigated. In vitro pull-down experiments showed that at a concentration of 100µM, the newly synthesized compounds had varying degrees of inhibitory effects on CRM1 binding to NES in vitro ( Figure 5 A), among which compounds 1, 2, 8 and 9 had the strongest inhibitory effects, followed by compound 3. After the concentration was reduced to 20µM, the inhibitory effects of all compounds weakened, and compounds 1, 2, 3, 8 and 9 still had significant inhibitory effects ( Figure 5 B). Immunofluorescence experiments were used to observe the phenomenon and found that at a concentration of 100 μM, compounds 2-8 all showed enhanced nuclear fluorescence, among which compounds 3-5 and compound 8 had the strongest inhibitory effect (see Figure 6 When the concentration dropped to 50µM, compounds 2-8 were still found to have a significant inhibitory effect on CRM1, as confirmed by the ratio of NES fluorescence intensity in the nucleus. However, the statistical results showed that the ability to inhibit CRM1 weakened as the concentration of the compound decreased, with compound 8 still showing a strong inhibitory effect on CRM1 (see Figure 7 To further understand the inhibitory effect of the compounds on CRM11-mediated nuclear export, the concentration of compounds 2-8 was reduced to 20µM for experiments. The results showed that, except for compounds 3 and 8, the inhibitory effects of the other compounds on CRM1 were significantly weakened. Compound 8 still maintained a high inhibitory effect on CRM1. Although the inhibitory effect of compound 3 was also weakened, its inhibitory effect was still relatively significant (see Figure 8 ).

[0029] Example 3: Inhibitory effect of compounds 1-9 on tumor cell growth Compounds 1-9 were studied for their anti-tumor cell growth effects. Figure 9 It can be seen that at a concentration of 100µM, compounds 1, 3, 5, 8 and 9 have a strong inhibitory effect on the growth of Hela cells, while at a concentration of 20µM, the inhibitory ability of these compounds weakened to varying degrees, among which compound 3 had the strongest inhibitory effect on the growth of Hela cells. Figure 10 It can be seen that compounds 1, 3, 8 and 9 have strong inhibitory effects on the growth of A549 cells (similar to Hela cells), among which compound 3 has the strongest inhibitory effect.

[0030] These results demonstrate that compounds 1-9 significantly enhance the ability to inhibit CRM1 nuclear export complex formation in vivo. In particular, compounds 3 and 8 inhibited CRM1 nuclear export complex formation in vitro, significantly inhibited CRM1-mediated cargo nuclear export in vivo, and inhibited the growth of Hela and A549 cells. Therefore, they can be used to prepare broad-spectrum anti-tumor non-covalent CRM1 inhibitors.

[0031] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, improvements, etc. made within the spirit and principles of the present invention should be included in the scope of protection of the present invention.

Claims

1. A small molecule compound, characterized in that The molecular structure of the small molecule compound is shown in formula (I) or formula (II): （Ⅰ） （Ⅱ）。 2. Use of the small molecule compound according to claim 1 in the preparation of a non-covalent CRM1 inhibitor and / or a medicament for preventing and / or treating a CRM1-related disease.

3. The use according to claim 2, characterized in that The diseases associated with CRM1 include pancreatic cancer, endometrial cancer, colorectal cancer, breast cancer, ovarian cancer, lung cancer and cervical cancer.

4. A non-covalent CRM1 inhibitor, characterized in that The non-covalent CRM1 inhibitor comprises at least one of the small molecule compounds of claim 1 .

5. A broad-spectrum anti-tumor drug, characterized in that: The broad-spectrum anti-tumor drug is a preparation prepared with the small molecule compound according to claim 1 or the non-covalent CRM1 inhibitor according to claim 4 as an active ingredient, and pharmaceutically acceptable excipients or auxiliary ingredients.

6. The broad-spectrum anti-tumor drug according to claim 5, characterized in that The broad-spectrum anti-tumor drug is administered orally, by injection, by rectal perfusion, by sublingual administration, by inhalation, by topical administration in vitro, or by systemic transdermal administration.

7. The broad-spectrum anti-tumor drug according to claim 6, characterized in that The dosage forms of the broad-spectrum anti-tumor drug include injections, capsules, tablets, sustained-release preparations, suppositories and sprays.

Images