A hsp27 inhibitor and methods of making and pharmaceutical uses thereof

Abstract

The present application provides a Hsp27 inhibitor, its preparation method and pharmaceutical application. The Hsp27 inhibitor is a compound shown in structural formula I or its pharmaceutically acceptable salt: wherein n=1, 2, 3 or 4; the Hsp27 inhibitor is used for treating diseases related to Hsp27 expression amount such as cancer.

Description

TECHNICAL FIELD

[0001] The present application belongs to the technical field of chemical biology, pharmacy and the like, and particularly relates to an Hsp27 inhibitor, a preparation method thereof and pharmaceutical application thereof. BACKGROUND

[0002] At present, chemotherapy and surgery are still the preferred choices for most early cancer patients. However, the side effects of chemotherapy are relatively high and have a greater impact on patients. In addition, cancer cells are prone to drug resistance during the current chemotherapy treatment process, leading to cancer recurrence and ineffective prognosis. Therefore, the current new anticancer drugs urgently need low side effects, effective reduction of drug resistance and specific targeting of cancer cells.

[0003] Heat shock protein 27 (Hsp27) is a small heat shock protein widely present in cells, which can regulate the balance between cell survival and death to cope with different stress conditions. In order to survive in the microenvironment of hypoxia and ischemia, Hsp27 is overexpressed in many cancer cells, such as liver cancer cells, lung cancer cells, gastrointestinal cancer cells, breast cancer cells, ovarian cancer cells and prostate cancer cells, etc. Previous studies have shown that Hsp27 regulates the occurrence, development, metastasis and drug resistance of various cancers. Studies have also shown that Hsp27 affects the migration and invasion of cancer cells through the NF-κB pathway, and also affects the growth of cancer cells. Another study has shown that overexpression of Hsp27 promotes TGFβ-mediated activity of matrix metalloproteinase 2 (MMP-2), thereby promoting cancer cell metastasis. In addition, Hsp27 also regulates cell aging or apoptosis by controlling the p53 pathway. Hsp27 plays an important role in the metastasis and drug resistance of cancer cells, and therefore Hsp27 is considered to be one of the cancer treatment targets with great significance. However, due to the lack of ATP binding sites, Hsp27 is difficult to be targeted by small molecule drugs. Previously, RP101 (broufadin) and Zerun bone (ZER) were considered to be effective inhibitors of Hsp27, and had certain effects in limiting the growth of cancer and prolonging the life of patients with advanced pancreatic cancer. However, the known Hsp27 inhibitors have low specificity and strong toxic side effects, which cannot meet the clinical application. In view of the foregoing, there is still much room for improvement for Hsp27 inhibitors.

[0004] Previously, Academician Kong Xiangluo et al. isolated various active ingredients from the traditional Chinese medicine gamboge. Among them, they found that 1,3,5-trihydroxy-13,13-dimethyl-2H-pyrano[7,6-b]anthracenone (TDP) can inhibit Hsp27 and mediate apoptosis of hepatocellular carcinoma cells 5 . The maximum half inhibitory concentration (IC 50) is 8 μM. Although daily administration of 20 mM TDP can effectively inhibit the growth of tumors in mice, it is difficult to be applied in clinical use due to its low bioavailability, poor cancer cell selectivity and large dosage used. SUMMARY

[0005] The present application develops a novel structure, good efficacy and high selectivity Hsp27 inhibitor for treating Hsp27 related diseases such as cancer.

[0006] The present application provides a compound or a pharmaceutically acceptable salt thereof, wherein the compound has a structure shown in structural formula I:

[0007]

[0008] wherein n = 1, 2, 3 or 4.

[0009] According to a specific embodiment of the present application, preferably, the compound has a structure shown in structural formula II:

[0010]

[0011] Some embodiments of the present application relate to a compound shown in structural formula II or a pharmaceutically acceptable salt thereof, wherein the compound is selected from one of the following structures:

[0012]

[0013]

[0014] According to a specific embodiment of the present application, preferably, the compound has a structure shown in structural formula III:

[0015]

[0016] Some embodiments of the present application relate to a compound shown in structural formula III or a pharmaceutically acceptable salt thereof, wherein the compound is selected from one of the following structures:

[0017]

[0018] The present application also provides a preparation method of the above-mentioned compound, which comprises the following steps:

[0019] (1) reacting fluorinated-2-hydroxybenzoic acid and m-phloroglucinol or o-phloroglucinol to obtain a compound shown in structural formula IV;

[0020] (2) reacting the compound shown in structural formula IV with 3-methyl-2-butenal to obtain a compound shown in structural formula I;

[0021] wherein the compound shown in structural formula IV:

[0022]

[0023] According to the specific embodiment of the present application, preferably, the preparation method of the compound comprises the following steps:

[0024] (1) dissolving fluorine-2-hydroxybenzoic acid and phloroglucinol or phloroglucinol in Eaton reagent at a molar ratio of 0.5-2, stirring at 50-150°C for 0.5-6 hours to obtain a compound shown in structural formula IV;

[0025] (2) dissolving the compound shown in structural formula IV and Ca(OH)2 in methanol at a molar ratio of 0.3-3, then adding 3-methyl-2-butenal, stirring at room temperature for 3 days to obtain a compound shown in structural formula I, wherein the molar ratio of 3-methyl-2-butenal to the compound shown in structural formula IV is 0.5-5.

[0026] According to the specific embodiment of the present application, preferably, in the above preparation method, the molar ratio of fluorine-2-hydroxybenzoic acid and phloroglucinol or phloroglucinol is 0.5-2.5.

[0027] According to the specific embodiment of the present application, preferably, in the above preparation method, the molar ratio of the compound shown in structural formula IV to Ca(OH)2 is 1:2.

[0028] According to the specific embodiment of the present application, taking HK-3 as an example, the preparation method of the above compound comprises the following steps:

[0029] (1) using 5-fluoro-2-hydroxybenzoic acid and phloroglucinol as raw materials to react to obtain a compound shown in structural formula V;

[0030] (2) reacting the compound shown in structural formula V with 3-methyl-2-butenal to obtain compound HK-3;

[0031]

[0032] According to the specific embodiment of the present application, taking HK-3 as an example, the above compound is prepared by the following specific steps:

[0033] (1) dissolving 5-fluoro-2-hydroxybenzoic acid (1 molar equivalent) and phloroglucinol (1 molar equivalent) in Eaton reagent, stirring the reaction mixture at 100°C for 0.5 hours; after the reaction is completed, cooling to room temperature, pouring the reaction mixture into ice and stirring for 2 hours to form a thin slurry; collecting the solid after filtration and washing with water; then purifying by silica gel chromatography (purification conditions: petroleum ether / ethyl acetate = 2:1, V / V) to obtain a light yellow solid (a compound shown in structural formula V);

[0034] (2) The light yellow solid (1 mole equivalent) and Ca(OH)2(2 mole equivalents) were mixed with methanol under stirring, 3-methyl-2-butenal (5 mole equivalents) was added to the mixture, after stirring at room temperature for 3 days, methanol was removed under vacuum, the reaction mixture was diluted with ethyl acetate, then the organic layer was washed with 2N HCl, water and brine, dried over Na2SO4, and the solvent was removed under vacuum, and purified by silica gel column chromatography (purification condition: ethyl acetate / petroleum ether = 1:99) to obtain HK-3 in the form of a yellow solid.

[0035] The present application also provides a compound represented by structural formula IV:

[0036]

[0037] The present application also provides a pharmaceutical composition comprising the above-mentioned compound or a pharmaceutically acceptable salt thereof as an active ingredient, and one or more pharmaceutically acceptable carriers or excipients.

[0038] The present application also provides the use of the above-mentioned compound or a pharmaceutically acceptable salt thereof or the above-mentioned pharmaceutical composition in the preparation of a medicament for treating a disease associated with the expression level of heat shock protein 27.

[0039] The present application also provides the use of the above-mentioned compound or a pharmaceutically acceptable salt thereof or the above-mentioned pharmaceutical composition in the preparation of a medicament for treating a disease associated with the inhibition of the expression of heat shock protein 27.

[0040] According to a specific embodiment of the present application, preferably, in the above-mentioned use, the disease is selected from cancer.

[0041] According to a specific embodiment of the present application, preferably, the cancer is selected from liver cancer, lung cancer, gastric cancer, gastrointestinal cancer or glioblastoma.

[0042] Unless otherwise stated, the terms used in the specification and claims have the following meanings:

[0043] "Pharmaceutically acceptable salt" or "a pharmaceutically acceptable salt thereof" refers to a salt of a compound of the present application which retains the biological effectiveness and properties of the free acid or the free base, and which is obtained by reaction of the free acid with a non-toxic inorganic or organic base, or the free base with a non-toxic inorganic or organic acid.

[0044] "Pharmaceutical composition" refers to a mixture of one or more compounds of the present application, a pharmaceutically acceptable salt or prodrug thereof, and other chemical components, such as pharmaceutically acceptable carriers, excipients, and / or one or more other therapeutic or active agents.

[0045] "Carrier" means a material that does not cause significant irritation to an organism and does not abrogate the biological activity and properties of the administered compound.

[0046] "Excipient" means an inert substance added to a pharmaceutical composition to facilitate administration of a compound. Non-limiting examples include calcium carbonate, calcium phosphate, sugars, starches, cellulose derivatives (including microcrystalline cellulose), gelatin, vegetable oils, polyethylene glycols, diluents, granulating agents, lubricants, binders, and disintegrating agents.

[0047] The HK-3 of the present application can effectively inhibit the expression of Hsp27 and promote the apoptosis of various cancers, and has more advantages than the original product TDP isolated from Garcinia cambogia in inhibiting liver cancer cells; the selectivity of HK-3 to cancer cells is stronger than that of TDP; and the binding force of HK-3 to Hsp27 protein is higher than that of TDP to Hsp27 protein. The Hsp27 inhibitor HK-3 of the present application can effectively inhibit the growth, migration and invasion of various cancers, and can be used as a substitute or auxiliary drug for anticancer drugs in a clinical environment. BRIEF DESCRIPTION OF DRAWINGS

[0048] Figure 1A is a micro-thermal mobility detection result graph;

[0049] Figure 1B is an immunofluorescence localization graph of Hsp27 of HepG2 cells treated by HK-3;

[0050] Figure 1C is an immunoblotting result graph of HepG2 cell lysate treated by HK-3;

[0051] Figure 1D is a fluorescence quantitative analysis result graph of HepG2 cell lysate treated by HK-3;

[0052] Figure 2A is a morphology graph of HepG2 cells after treatment;

[0053] Figure 2B is a survival rate result graph of HepG2 cells after treatment;

[0054] Figure 2C is a morphology graph of normal liver cells after treatment;

[0055] Figure 2D is a survival rate result graph of normal liver cells after treatment;

[0056] Figure 3A is a protein expression result graph of apoptosis-related factors in HepG2 cells;

[0057] Figure 3BStatistical graph of mRNA transcription level of apoptosis-related factors in HepG2 cells;

[0058] Figure 4A Morphology graph of various cancer cells after HK-3 treatment;

[0059] Figure 4B Survival rate result graph of various cancer cells after HK-3 treatment;

[0060] Figure 5 Result graph of HK-3 inhibiting tumor cell growth and migration;

[0061] Figure 6 Result graph of HK-3 inhibiting tumor cell migration and infiltration. DETAILED DESCRIPTION

[0062] In order to have a clearer understanding of the technical features, objectives and beneficial effects of the present application, the technical solutions of the present application are described in detail below, but it should not be understood as limiting the scope of the present application.

[0063] Example 1

[0064] This example provides a compound HK-3, which is prepared as follows:

[0065] (1) 5-Fluoro-2-hydroxybenzoic acid (1.56 g, 10.0 mmol) and m-benzene triol (1.26 g, 10.0 mmol) were dissolved in Eaton's reagent (25.0 mL); the reaction mixture was stirred at 100°C for 0.5 h; after the reaction was completed, it was cooled to room temperature, the reaction mixture was poured into ice and stirred for 2 h to form a thin slurry; the solid was collected after filtration and washed with water; then purified by silica gel chromatography (purification condition: petroleum ether / ethyl acetate = 2:1, V / V) to obtain a light yellow solid compound (1.47 g, 60%);

[0066] (2) The light yellow solid compound (250 mg, 1.02 mmol) and Ca(OH)2(150 mg, 2.05 mmol) were stirred and mixed with methanol (30 ml), 3-methyl-2-butenal (0.5 mL, 5.12 mmol) was added to the mixture, after stirring at room temperature for 3 days, the methanol was removed under vacuum, the reaction mixture was diluted with ethyl acetate (30 mL), then the organic layer was washed with 2N HCl, water and brine, dried over Na2SO4, and the solvent was removed under vacuum, and purified by silica gel column chromatography (purification condition: 1% ethyl acetate / petroleum ether) to obtain HK-3 in the form of a yellow solid (126 mg, 40%).

[0067] HRMS (ESI): m / z [M+H] + calcd. for [C18 H 14 O4F] + :313.0871, found:313.0870.

[0068] 1 H NMR (400 MHz, CDC13) δ: 12.96 (s, 1H), 7.86 (d, J = 5.8 Hz, 1H), 7.41 (s, 2H), 6.72 (d, J = 9.3 Hz, 1H), 6.33 (s, 1H), 5.61 (d, J = 9.5 Hz, 1H), 1.48 (s, 6H); 13 C NMR (101 MHz, CDC13) δ: 179.9, 161.1, 159.9, 157.5, 157.1, 152.1, 127.7, 122.8 (d, J = 25.1 Hz), 121.4 (d, J = 7.1 Hz), 119.5 (d, J = 7.8 Hz), 110.7 (d, J = 23.8 Hz), 104.7, 103.3, 95.1, 78.4, 28.4; 19 F NMR (376 MHz, CDC13) δ: -116.8.

[0069] The experiments and specific experimental methods involved in the test examples of the present application are as follows:

[0070] 1. Cell culture

[0071] The cell culture adopted in the present application involves human normal hepatocytes (L-O2), hepatoma cells (HepG2, Hep3B and Huh7), lung cancer cells (A649), gastrointestinal cancer cells (SW620), gastric cancer cells (MKN28), human glioblastoma cells (U251), rhabdomyosarcoma cells (RD), cervical cancer cells (Hela) and mouse breast cancer cells 4T1 cells, which are all purchased from the American Type Culture Collection (ATCC, Manassas, VA, USA); the culture method is as follows: Dulbecco's Modified Eagle Medium (DMEM) containing 10% fetal bovine serum (FBS) is used for cell culture, supplemented with 100 U / ml penicillin and 100 μg / ml streptomycin; all cells are cultured in a 5% CO2 incubator at 37°C.

[0072] 2. Western Blotting

[0073] Total protein was extracted from cell lysate in RIPA buffer with protease and phosphatase inhibitors (Roche); 10-30 pg of protein was loaded for SDS-PAGE in total and transferred to PVDF membrane; the membrane was blocked with Tris-buffered saline buffer containing 5% skim milk, 0.1% Tween-20 (TBST) for 1 hour at room temperature and incubated with primary antibody overnight at 4°C; after washing 3 times with TBST, the membrane was incubated with the corresponding horseradish peroxidase (HRP)-conjugated secondary antibody for 1 hour at room temperature; the band density was quantified using Quantity One software (Bio-Rad); GADPH was used as the internal control.

[0074] 3. RNA extraction and real-time polymerase chain reaction (qRT-PCR)

[0075] Total RNA was extracted from cell lysate using TRIzol reagent (Ambion, Life Technologies). Quantitative real-time PCR (qRT-PCR) was performed using SYBR Green Mix (Life Technologies) on an Applied Biosystems StepOne Real-Time PCR System. The relative expression of each gene was calculated compared to GADPH mRNA using the AACt method. Primer sequences are listed in Table 1.

[0076] Table 1 Primer sequences required for quantitative real-time PCR

[0077]

[0078] 4. Cell viability assay

[0079] HepG2 and other cancer cells were seeded into 96-well plates (5,000 cells / well) and treated with serial dilutions of TDP or HK-3 for 48 hours for cell viability assay. 10 mΐ of MTT was added to the cultured cells in 100 mΐ of culture medium and incubated at 37°C for 4 h. Subsequently, 100 mΐ of dimethyl sulfoxide was added. The optical density was measured at a wavelength of 570 nm.

[0080] 5. Microscale thermophoresis (MST) assay

[0081] Hsp27 recombinant protein with His-tag was mixed with Monolith TM RED dye and incubated at room temperature for half an hour to generate fluorescent signal; subsequently, 500 nM HK-3 was gradient diluted 16 times and mixed with fluorescent-labeled Hsp27 protein, avoiding light for 15 minutes; then, Monolith TMNT.115 capillary glass tube to pipette micro-volume mixture and place in Monolith for microscale thermophoresis (MST) detection; LED / excitation power set to 40%, MST power set to medium.

[0082] 6. Wound assay

[0083] Wound assay

[0084] 7. Transwell assay

[0085] Transwell system purchased from Thermo Fish (140656, PC pore size 8 pm). HK-3 (100 nM, HepG2; 400 nM, 4T1) was added to the upper chamber with blank medium, and medium containing 10% fetal bovine serum was added to the lower chamber; after 16 hours of incubation, the cells migrated or invaded to the lower chamber membrane were fixed with 4% paraformaldehyde and stained with 0.1% crystal violet, and three non-overlapping fields were randomly selected under an optical microscope for cell counting.

[0086] 8. Crystal violet viability assay

[0087] The cells were carefully washed with PBS and stained with 0.5% crystal violet formalin solution at room temperature for 30 minutes, then the cells were carefully washed with ddH2O, and three non-overlapping fields were randomly selected under an optical microscope for photography.

[0088] 9. Statistical analysis

[0089] The results are expressed as mean ± standard deviation (SD). The significance of the difference between data was compared by two-tailed t-test, and P value less than 0.05 was defined as significant.

[0090] Test Example 1

[0091] This test example investigates the affinity of HK-3 to Hsp27 and the effect of HK-3 on Hsp27 expression.

[0092] The results of microscale thermophoresis (MST) analysis are shown in Table 1. Figure 1AAs shown, the Kd value of TDP to Hsp27 is 459 nM, but its signal / background noise ratio is 2.9, which is not effective binding, and the Kd value of HK-3 to Hsp27 is 214 nM, and its signal / background noise ratio is 11.0, which is effective binding. HK-3 (Kd = 214 nM, signal to noise ratio = 11.0) binds to Hsp27 much more effectively than TDP (459 nM).

[0093] Further investigation of the expression of Hsp27 in HepG2 cells after treatment with low concentration of HK-3 for 0, 24, 48 hours, by immunofluorescence staining, the results are shown in Figure 1B , it can be seen that the expression of Hsp27 is inhibited by HK-3; in order to further verify this guess, qPCR technology and Western blotting method were used to determine the transcription (results are shown in Figure 1D ) and translation (results are shown in Figure 1C ) levels of Hsp27 in HepG2 cells treated with HK-3 for 24 hours, and the results showed that HK-3 can effectively inhibit the expression of Hsp27.

[0094] Test Example 2

[0095] This test example investigates the inhibitory effect of HK-3 on liver cancer cells and the selectivity of HK-3 on cancer cells.

[0096] Comparison of the cytopathic effect (CPE) (cell morphology results are shown in Figure 2A ) and survival rate (results are shown in Figure 2B ) of HepG2 cells treated with HK-3 or TDP for 48 hours, the results showed that HK-3 has a significant inhibitory effect on liver cancer cells HepG2 (IC 50 = 13.79 nM); at the same time, the morphology (results are shown in Figure 2C ) and survival rate of normal liver cells (L-O2) treated with HK-3 or TDP for 48 hours were investigated, it can be seen that the half inhibitory concentration of HK-3 on normal liver cells (L-O2) is 388.40 nM (results are shown in Figure 2D ), and that of TDP is 12.45 μM (results are shown in Figure 2D ). The ratio of the half inhibitory concentration of liver cancer cells to that of normal liver cells was calculated, which was 28.17 for HK-3 and 1.96 for TDP. Therefore, HK-3 has high selectivity for cancer cells (the ratio of half inhibitory concentration is more than 10 times greater than that of TDP). In summary, HK-3 not only has a high inhibitory effect on liver cancer cells, but also has high selectivity, and has the potential to specifically remove cancer cells.

[0097] Test Example 3

[0098] This test example verifies the mechanism of HK-3 in inhibiting cancer cells.

[0099] Previous reports have shown that Hsp27 can hinder the response of cancer cells to apoptosis by inhibiting the expression of Caspase 3. Therefore, this test example verifies whether HK-3 can re-activate the response of cancer cells to apoptotic signals, thereby causing cancer cell death.

[0100] After HepG2 cells were treated with HK-3 or control solution for 48 hours, cell lysates were collected by RIPA buffer or TRISOL for further analysis; the results of Western blot analysis of cell extracts (see Figure 3A ) and real-time polymerase chain reaction (qPCR) (see Figure 3B ) showed that the apoptosis regulatory factor p53 and the main cell apoptosis executor caspase 3 in the cells were promoted after treatment with HK-3; at the same time, the activity of NF-κB, a related factor mediating tumor formation, was also inhibited by HK-3, so it can be considered that HK-3 induces apoptosis of hepatocarcinoma cells by inhibiting the expression level of Hsp27.

[0101] Test Example 4

[0102] This test example investigates the inhibitory effect of HK-3 on various cancer cells.

[0103] Since Hsp27 is highly expressed in various cancers, we believe that HK-3 can also act on other cancer cells. In order to verify this assumption, the inhibitory efficiency of HK-3 on several cancer cell lines (e.g. lung cancer cells (A549), gastrointestinal cancer cells (SW620), gastric cancer cells (MKN28), human glioblastoma cells (U251)) was further determined, and the results were consistent with the high inhibitory efficiency in hepatocarcinoma cell lines. HK-3 still has a strong inhibitory effect in different cancer cell lines (results see Figure 4A and Figure 4B ).

[0104] Test Example 5

[0105] This test example investigates the inhibitory effect of HK-3 on tumor cell growth and migration.

[0106] It is proved by scratch test that HK-3 inhibits the growth and migration of hepatocarcinoma cells HepG2, as shown in Figure 5 .

[0107] Test Example 6

[0108] This test example investigates the inhibitory effect of HK-3 on tumor cell migration and invasion.

[0109] The tumor cell (Hep3B) migration and tumor cell infiltration experiments prove that HK-3 inhibits the migration and infiltration of liver cancer cells, and the results are shown in Figure 6 .

[0110] The HK-3 of the present application can effectively inhibit the expression of Hsp27 and promote the apoptosis of various cancers, and the inhibitory effect on liver cancer cells can reach the nanomolar level (IC 50 = 13.79 nM), which is more advantageous than TDP (IC 50 = 8.0 μM) and the activity is increased by 580 times; the selectivity of HK-3 to cancer cells is 14 times that of TDP; and the binding force Kd value of HK-3 to Hsp27 protein is 214 nM, which is 2 times that of TDP.

[0111] The above data shows that HK-3 is an optimized and effective Hsp27 inhibitor, and the high activity, high selectivity and effective inhibition of various cancer cells make HK-3 expected to become an anti-cancer drug for clinical application.

Claims

1. A compound, or a pharmaceutically acceptable salt thereof, wherein, The compound has a structure shown in structural formula II: Formula II, wherein n = 1, 2, 3, or 4.

2. The compound or pharmaceutically acceptable salt thereof of claim 1, wherein, The compound shown in structural formula II is selected from one of the following structures: , , , , , , , , , , , , or .

3. A method for preparing the compound of claim 1 or 2, comprising the following steps: (1) reacting fluorinated-2-hydroxybenzoic acid and phloroglucinol to obtain a compound shown in structural formula VI; (2) reacting the compound shown in structural formula VI with 3-methyl-2-butenal to obtain the compound shown in structural formula II; wherein The compound shown in structural formula VI: Formula VI.

4. The method of claim 3, comprising the following steps: (1) dissolving fluorinated-2-hydroxybenzoic acid and phloroglucinol in an Eaton reagent at a molar ratio of 0.5-2, and stirring at a temperature of 50-150°C for 0.5-20 hours to obtain the compound shown in structural formula VI; (2) dissolving the compound shown in structural formula VI and Ca(OH)2 in methanol at a molar ratio of 0.3-3, and then adding 3-methyl-2-butenal, and stirring at room temperature to obtain the compound shown in structural formula II, wherein the molar ratio of 3-methyl-2-butenal to the compound shown in structural formula VI is 0.5-5.

5. The production method according to claim 4, wherein The molar ratio of the fluorinated-2-hydroxybenzoic acid to the phloroglucinol is 0.5-2.

5.

6. The production method according to claim 4, wherein The molar ratio of the compound shown in structural formula VI to Ca(OH)2 is 1:

2.

7. A pharmaceutical composition comprising the compound of claim 1 or 2 or a pharmaceutically acceptable salt thereof as an active ingredient, and one or more pharmaceutically acceptable carriers or excipients.

8. Use of the compound of claim 1 or 2 or a pharmaceutically acceptable salt thereof or the pharmaceutical composition of claim 7 in the preparation of a medicament for treating liver cancer, lung cancer, gastric cancer, gastrointestinal cancer, or glioblastoma.

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